U.S. patent application number 11/312812 was filed with the patent office on 2007-03-15 for inhibitors of protein arginine methyl transferases.
Invention is credited to Zhong Chen, Ashok Vinayak Purandare.
Application Number | 20070060589 11/312812 |
Document ID | / |
Family ID | 36602287 |
Filed Date | 2007-03-15 |
United States Patent
Application |
20070060589 |
Kind Code |
A1 |
Purandare; Ashok Vinayak ;
et al. |
March 15, 2007 |
Inhibitors of protein arginine methyl transferases
Abstract
A compound of formula I, or a stereoisomer, a tautomer, a
pharmaceutically acceptable salt or solvate thereof, ##STR1##
methods of using such compounds in the treatment of
hyperproliferative, inflammatory, infectious, and immunoregulatory
disorders and diseases; and to pharmaceutical compositions
containing such compounds.
Inventors: |
Purandare; Ashok Vinayak;
(Pennington, NJ) ; Chen; Zhong; (Princeton,
NJ) |
Correspondence
Address: |
LOUIS J. WILLE;BRISTOL-MYERS SQUIBB COMPANY
PATENT DEPARTMENT
P O BOX 4000
PRINCETON
NJ
08543-4000
US
|
Family ID: |
36602287 |
Appl. No.: |
11/312812 |
Filed: |
December 20, 2005 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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60637893 |
Dec 21, 2004 |
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Current U.S.
Class: |
514/252.05 ;
514/255.05; 514/256; 514/310; 514/318; 514/326; 544/238; 544/333;
544/405; 546/146; 546/194; 546/209 |
Current CPC
Class: |
C07C 237/42 20130101;
C07D 413/12 20130101; C07D 231/14 20130101; C07D 413/04 20130101;
C07D 409/12 20130101; C07D 417/14 20130101; C07D 403/12 20130101;
C07D 405/12 20130101; C07D 401/12 20130101; C07D 471/04 20130101;
C07D 231/38 20130101; C07D 417/12 20130101; C07D 413/14
20130101 |
Class at
Publication: |
514/252.05 ;
514/310; 514/326; 514/255.05; 514/256; 514/318; 544/238; 544/333;
544/405; 546/146; 546/194; 546/209 |
International
Class: |
A61K 31/506 20060101
A61K031/506; A61K 31/501 20060101 A61K031/501; A61K 31/497 20060101
A61K031/497; A61K 31/4709 20060101 A61K031/4709; A61K 31/4545
20060101 A61K031/4545; A61K 31/454 20060101 A61K031/454; C07D
417/02 20060101 C07D417/02; C07D 413/02 20060101 C07D413/02; C07D
403/02 20060101 C07D403/02 |
Claims
1. A compound of formula I, a stereoisomer, a tautomer, or a
pharmaceutically acceptable salt thereof, ##STR217## wherein: Ring
Q is phenyl; 5-membered heteroaryl, in which Z.sub.4 is a bond,
Z.sub.1, Z.sub.2, Z.sub.3 and Z.sub.5 are each independently C, N,
O, or S, and at least one of Z.sub.1, Z.sub.2, Z.sub.3 and Z.sub.5
is a heteroatom selected from N, O and S; or 6-membered heteroaryl,
in which Z.sub.1, Z.sub.2, Z.sub.3, Z.sub.4 and Z.sub.5 are each
independently C, or N, and at least one of Z.sub.1, Z.sub.2,
Z.sub.3, Z.sub.4 and Z.sub.5 is N; W is --C(.dbd.O)NR.sub.8--,
--NR.sub.9C(.dbd.O)--, --NR.sub.9C(.dbd.O)NR.sub.9--, ##STR218##
alternatively, W--(CH.sub.2)--(O)n-R.sub.7 is ##STR219## R.sub.1 is
H, halogen, CN, alkyl or substituted alkyl, O--C.sub.1-C.sub.4
alkyl, S--C.sub.1-C.sub.4 alkyl, or SO.sub.2--C.sub.1-C.sub.4
alkyl; R.sub.2 is H or C.sub.1-C.sub.4 alkyl; R.sub.3 is H, Me or
Et, or optionally R.sub.3 together with R.sub.4 may form a 5- or
6-membered heterocycle R.sub.4 is H, Me, Et, iso-propyl,
CH.sub.2Ph, OH, or OPh, or optionally R.sub.4 together with R.sub.3
may form a 5- or 6-membered heterocycle; R.sub.5 is nil, H, Me, Et,
propyl, iso-propyl, OMe, OEt, SMe, SO.sub.2Me, CF.sub.3, or
OCF.sub.3; R.sub.6 is nil, H, Me, or Et, or optionally R.sub.6
together with R.sub.8 may form a 5- or 6-membered heterocycle;
R.sub.7 is cycloalkyl or substituted cycloalkyl, heterocycle or
substituted heterocycle, or aryl or substituted aryl; R.sub.8 is H,
or Me, or optionally R.sub.8 together with R.sub.6 may form a 5- or
6-membered heterocycle; or alternatively R.sub.8 together with
R.sub.7 may form a 5- or 6-membered heterocycle or substituted
heterocycle; R.sub.9 is H, or Me; R.sub.10 is hydrogen, halogen,
haloalkyl, cyano, nitro, alkyl, cycloalkyl, alkenyl, cycloalkenyl,
alkynyl, heterocycle, aryl, OR, SR, S(.dbd.O)R, S(.dbd.O).sub.2R,
P(.dbd.O).sub.2R, S(.dbd.O).sub.2OR, P(.dbd.O).sub.2OR, NRR,
NRS(.dbd.O).sub.2R, NRP(.dbd.O).sub.2R, S(.dbd.O).sub.2NRR,
P(.dbd.O).sub.2NRR, C(.dbd.O)OR, C(.dbd.O)R, C(.dbd.O)NRR,
OC(.dbd.O)R, OC(.dbd.O)NRR, NRC(.dbd.O)OR, NRC(.dbd.O)NRR,
NRS(.dbd.O).sub.2NRR, NRP(.dbd.O).sub.2NRR,
NR.sub.bC(.dbd.O)R.sub.a, or NRP(.dbd.O).sub.2R, wherein R is
independently hydrogen, alkyl, cycloalkyl, alkenyl, cycloalkenyl,
alkynyl, heterocycle, or aryl; m is 0, 1, 2 or 3; n is 0 or 1; and
p is 1, 2 or 3.
2. The compound of claim 1, wherein ring Q is 5-membered
heteroaryl, in which Z.sub.4 is a bond, Z.sub.1, Z.sub.2, Z.sub.3
and Z.sub.5 are each independently C, N, O, or S, and at least one
of Z.sub.1, Z.sub.2, Z.sub.3 and Z.sub.5 is a heteroatom selected
from N, O and S; or 6-membered heteroaryl, in which Z.sub.1,
Z.sub.2, Z.sub.3, Z.sub.4 and Z.sub.5 are each independently C, or
N, and at least one of Z.sub.1, Z.sub.2, Z.sub.3, Z.sub.4 and
Z.sub.5 is N.
3. The compound of claim 2, wherein ring Q is 5-membered
heteroaryl, in which Z.sub.4 is a bond, Z.sub.1, Z.sub.2, Z.sub.3
and Z.sub.5 are each independently C, N, O, or S, and at least one
of Z.sub.1, Z.sub.2, Z.sub.3 and Z.sub.5 is a heteroatom selected
from N, O and S.
4. The compound of claim 3, wherein Z.sub.4 is a bond, Z.sub.1 and
Z.sub.2, are each independently N; Z.sub.3 and Z.sub.5 are each
independently C.
5. The compound of claim 4, wherein R.sub.7 is aryl or substituted
aryl.
6. The compound of claim 5, wherein R.sub.7 is phenyl or
substituted phenyl.
7. The compound of claim 1, wherein W is --C(.dbd.O)NR.sub.8--.
8. The compound of claim 7, wherein ring Q is 5-membered
heteroaryl, in which Z.sub.4 is a bond, Z.sub.1, Z.sub.2, Z.sub.3
and Z.sub.5 are each independently C, N, O, or S, and at least one
of Z.sub.1, Z.sub.2, Z.sub.3 and Z.sub.5 is a heteroatom selected
from N, O and S.
9. The compound of claim 8, wherein Z.sub.4 is a bond, Z.sub.1 and
Z.sub.2, are each independently N; Z.sub.3 and Z.sub.5 are each
independently C.
10. The compound of claim 9, wherein R.sub.7 is aryl or substituted
aryl.
11. The compound of claim 10, wherein R.sub.7 is phenyl or
substituted phenyl.
12. The compound of claim 1, wherein W is ##STR220##
13. The compound of claim 12, wherein ring Q is 5-membered
heteroaryl, in which Z.sub.4 is a bond, Z.sub.1, Z.sub.2, Z.sub.3
and Z.sub.5 are each independently C, N, O, or S, and at least one
of Z.sub.1, Z.sub.2, Z.sub.3 and Z.sub.5 is a heteroatom selected
from N, O and S.
14. The compound of claim 13, wherein Z.sub.4 is a bond, Z.sub.1
and Z.sub.2, are each independently N; Z.sub.3 and Z.sub.5 are each
independently C.
15. The compound of claim 14, wherein R.sub.7 is aryl or
substituted aryl.
16. The compound of claim 15, wherein R.sub.7 is phenyl or
substituted phenyl.
17. The compound of claim 1, wherein R.sub.2 is H, R.sub.3 is H,
R.sub.4 is Me, Et, OH, or Ph.
18. The compound of claim 17, wherein R.sub.4 is Me.
19. The compound of claim 18, wherein R.sub.4 is (S--) Me.
20. The compound of claim 19, wherein ring Q is 5-membered
heteroaryl, in which Z.sub.4 is a bond, Z.sub.1, Z.sub.2, Z.sub.3
and Z.sub.5 are each independently C, N, O, or S, and at least one
of Z.sub.1, Z.sub.2, Z.sub.3 and Z.sub.5 is a heteroatom selected
from N, O and S.
21. The compound of claim 20, wherein Z.sub.4 is a bond, Z.sub.1
and Z.sub.2, are each independently N; Z.sub.3 and Z.sub.5 are each
independently C.
22. The compound of claim 19, wherein W is
--C(.dbd.O)NR.sub.8--.
23. The compound of claim 22, wherein ring Q is 5-membered
heteroaryl, in which Z.sub.4 is a bond, Z.sub.1, Z.sub.2, Z.sub.3
and Z.sub.5 are each independently C, N, O, or S, and at least one
of Z.sub.1, Z.sub.2, Z.sub.3 and Z.sub.5 is a heteroatom selected
from N, O and S.
24. The compound of claim 23, wherein Z.sub.4 is a bond, Z.sub.1
and Z.sub.2, are each independently N; Z.sub.3 and Z.sub.5 are each
independently C.
25. The compound of claim 19, wherein W is ##STR221##
26. The compound of claim 25, wherein ring Q is 5-membered
heteroaryl, in which Z.sub.4 is a bond, Z.sub.1, Z.sub.2, Z.sub.3
and Z.sub.5 are each independently C, N, O, or S, and at least one
of Z.sub.1, Z.sub.2, Z.sub.3 and Z.sub.5 is a heteroatom selected
from N, O and S.
27. The compound of claim 26, wherein Z.sub.4 is a bond, Z.sub.1
and Z.sub.2, are each independently N; Z.sub.3 and Z.sub.5 are each
independently C.
28. A pharmaceutical composition comprising at least one compound
according to claim 1 and a pharmaceutically-acceptable carrier or
diluent.
29. A pharmaceutical composition of claim 28, further comprising at
least one other anti-cancer agent or cytotoxic agent.
30. The pharmaceutical composition of claim 29, wherein said
anti-cancer or cytotoxic agent is selected from the group
consisting of tamoxifen, toremifene, raloxifene, droloxifene,
iodoxifene, megestrol acetate, anastrozole, letrozole, borazole,
exemestane, flutamide, nilutamide, bicalutamide, cyproterone
acetate, gosereline acetate, leuprolide, finasteride,
metalloproteinase inhibitors, inhibitors of urokinase plasminogen
activator receptor function, growth factor antibodies, growth
factor receptor antibodies, bevacizumab, cetuximab, tyrosine kinase
inhibitors, serine/threonine kinase inhibitors, methotrexate,
5-fluorouracil, purine and adenosine analogues, cytosine
arabinoside, doxorubicin, daunomycin, epirubicin, idarubicin,
mitomycin-C, dactinomycin, mithramycin, cisplatin, carboplatin,
nitrogen mustard, melphalan, chlorambucil, busulphan,
cyclophosphamide, ifosfamide, nitrosoureas, thiotepa, vincristine,
vinorelbine, vinblastine, vinflunine, paclitaxel, docetaxel,
epothilone analogs, discodermolide analogs, eleutherobin analogs,
etoposide, teniposide, amsacrine, topotecan, flavopyridols,
proteasome inhibitors including bortezomib and biological response
modifiers, androgen receptor antagonists, LH/RH antagonists, taxane
analogues, and estrogen receptor antagonists.
31. A method of inhibiting the activity of CARM-1 which comprises
administering to a mammalian species in need thereof an effective
amount of at least one compound of formula I: ##STR222## or a
stereoisomer, a tautomer, a pharmaceutically acceptable salt or
solvate thereof, wherein: Ring Q is phenyl; 5-membered heteroaryl,
in which Z.sub.4 is a bond, Z.sub.1, Z.sub.2, Z.sub.3 and Z.sub.5
are each independently C, N, O, or S, and at least one of Z.sub.1,
Z.sub.2, Z.sub.3 and Z.sub.5 is a heteroatom selected from N, O and
S; or 6-membered heteroaryl, in which Z.sub.1, Z.sub.2, Z.sub.3,
Z.sub.4 and Z.sub.5 are each independently C, or N, and at least
one of Z.sub.1, Z.sub.2, Z.sub.3, Z.sub.4 and Z.sub.5 is N; W is
--C(.dbd.O)NR.sub.8--, --NR.sub.9C(.dbd.O)--, ##STR223## R.sub.1 is
H, halogen, CN, alkyl or substituted alkyl, O--C.sub.1-C.sub.4
alkyl, S--C.sub.1-C.sub.4 alkyl, or SO.sub.2--C.sub.1-C.sub.4
alkyl; R.sub.2 is H or C.sub.1-C.sub.4 alkyl; R.sub.3 is H, Me or
Et, or optionally R.sub.3 together with R.sub.4 may form a 5- or
6-membered heterocycle R.sub.4 is H, Me, Et, iso-propyl,
CH.sub.2Ph, OH, or OPh, or optionally R.sub.4 together with R.sub.3
may form a 5- or 6-membered heterocycle; R.sub.5 is nil, H, Me, Et,
propyl, iso-propyl, OMe, OEt, SMe, SO.sub.2Me, CF.sub.3, or
OCF.sub.3; R.sub.6 is nil, H, Me, or Et, or optionally R.sub.6
together with R.sub.8 may form a 5- or 6-membered heterocycle;
R.sub.7 is cycloalkyl or substituted cycloalkyl, heterocycle or
substituted heterocycle, or aryl or substituted aryl; R.sub.8 is H,
or Me, or optionally R.sub.8 together with R.sub.6 may form a 5- or
6-membered heterocycle; or alternatively R.sub.8 together with
R.sub.7 may form a 5- or 6-membered heterocycle or substituted
heterocycle; R.sub.9 is H, or Me; m is 0, 1, 2 or 3; n is 0 or 1;
and p is 1, 2 or 3.
32. A method for treating a condition or disorder comprising
administering to a mammalian species in need thereof a
therapeutically effective amount of at least one compound of
formula I: ##STR224## or a stereoisomer, a tautomer, a
pharmaceutically acceptable salt or solvate thereof, wherein: Ring
Q is phenyl; 5-membered heteroaryl, in which Z.sub.4 is a bond,
Z.sub.1, Z.sub.2, Z.sub.3 and Z.sub.5 are each independently C, N,
O, or S, and at least one of Z.sub.1, Z.sub.2, Z.sub.3 and Z.sub.5
is a heteroatom selected from N, O and S; or 6-membered heteroaryl,
in which Z.sub.1, Z.sub.2, Z.sub.3, Z.sub.4 and Z.sub.5 are each
independently C, or N, and at least one of Z.sub.1, Z.sub.2,
Z.sub.3, Z.sub.4 and Z.sub.5 is N; W is --C(.dbd.O)NR.sub.8--,
--NR.sub.9C(.dbd.O)--, ##STR225## R.sub.1 is H, halogen, CN, alkyl
or substituted alkyl, O--C.sub.1-C.sub.4 alkyl, S--C.sub.1-C.sub.4
alkyl, or SO.sub.2--C.sub.1-C.sub.4 alkyl; R.sub.2 is H, or
C.sub.1-C.sub.4 alkyl; R.sub.3 is H, Me or Et, or optionally
R.sub.3 together with R.sub.4 may form a 5- or 6-membered
heterocycle R.sub.4 is H, Me, Et, iso-propyl, CH.sub.2Ph, OH, or
OPh, or optionally R.sub.4 together with R.sub.3 may form a 5- or
6-membered heterocycle; R.sub.5 is nil, H, Me, Et, propyl,
iso-propyl, OMe, OEt, SMe, SO.sub.2Me, CF.sub.3, or OCF.sub.3;
R.sub.6 is nil, H, Me, or Et, or optionally R.sub.6 together with
R.sub.8 may form a 5- or 6-membered heterocycle; R.sub.7 is
cycloalkyl or substituted cycloalkyl, heterocycle or substituted
heterocycle, or aryl or substituted aryl; R.sub.8 is H, or Me, or
optionally R.sub.8 together with R.sub.6 may form a 5- or
6-membered heterocycle; or alternatively R.sub.8 together with
R.sub.7 may form a 5- or 6-membered heterocycle or substituted
heterocycle; R.sub.9 is H, or Me; m is 0, 1, 2 or 3; n is 0 or 1;
and p is 1, 2 or 3; and wherein said condition or disorder is
selected from the group consisting of proliferate diseases,
cancers, benign prostate hypertrophia, benign prostatic
hyperplasia, adenomas and neoplasies of the prostate, benign or
malignant tumor cells containing the androgen receptor, brain
cancer, skin cancer, bladder cancer, lymphatic cancer, liver
cancer, kidney cancer, pancreatic cancer, prostate cancer,
hirsutism, acne, precocious puberty, angiogenic conditions or
disorders, hyperpilosity, inflammation, immune modulation,
seborrhea, endometriosis, polycystic ovary syndrome, androgenic
alopecia, hypogonadism, osteoporosis, suppressing spermatogenesis,
male and female sexual dysfunction, libido, cachexia, anorexia,
inhibition of muscular atrophy in ambulatory patients, androgen
supplementation for age related decreased testosterone levels in
men, cancers expressing the estrogen receptor, breast cancer,
ovarian cancer, uterine cancer, endometrial cancer, hot flushes,
vaginal dryness, menopause, amennoreahea, dysmennoreahea,
contraception, pregnancy termination, cancers containing the
progesterone receptor, cyclesynchrony, meniginoma, fibroids, labor
induction, autoimmune diseases, Alzheimer's disease, psychotic
disorders, drug dependence, non-insulin dependent Diabetes
Mellitus, dopamine receptor mediated disorders, heart disease,
congestive heart failure, disregulation of cholesterol homeostasis,
and attenuating the metabolism of a pharmaceutical agent.
33. A method for treating cancer comprising administering to a
patient in need thereof, a therapeutically effective amount of a
compound having the formula I according to claim 1, wherein the
cancer is breast cancer, lung cancer, ovarian cancer, prostate
cancer, leukemia, lymphoma, glioblastoma, brain cancer, melanoma,
or colon cancer.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] The present application claims the priority benefit of U.S.
Provisional Application No. 60/637,893, filed Dec. 21, 2004 which
is expressly incorporated fully herein by reference
FIELD OF THE INVENTION
[0002] The present invention relates to novel compounds which are
inhibitors of Protein ARginine Methyl Transferases (PRMTs), to
methods of using such compounds for inhibiting protein methyl
transferases in the treatment of hyperproliferative, inflammatory,
infectious, and immunoregulatory disorders and diseases, and to
pharmaceutical compositions containing such compounds.
[0003] The invention also encompasses pharmaceutical compositions
containing these compounds. The compounds and pharmaceutical
compositions of the invention are particularly well suited as
inhibitors of protein methyl transferases and, consequently, can be
advantageously used as therapeutic agents for the treatment of,
including cancer, asthma, COPD, and allergic diseases; rheumatoid
arthritis, atherosclerosis, and psoriasis; solid organ transplant
rejection, osteoarthritis, and inflammatory bowel syndrome. This
invention also relates to methods of using the compounds of this
invention alone or in combination with other pharmaceutically
active agents.
BACKGROUND OF THE INVENTION
[0004] PRMTs are enzymes that catalyze the transfer of methyl
groups from S-Adenosyl-L-Methionine (SAM) to specific arginine
residues of proteins. Arginine methylation of proteins has been
implicated to play roles in pre-mRNA splicing, nucleo-cytoplasmic
RNA transport, signal transduction and transcriptional activation.
To date seven family members have been identified (PRMTs 1-7) in
mammalian cells and they each appear to have distinct substrate
preferences. PRMT1 has been shown to methylate Histone H4 and this
results in activation of transcription. Coactivator Associated
Arginine Methyltransferase-I (CARM-1) (also called PRMT-4) has been
shown to methylate Histone H3 both in vitro and in vivo and it is
speculated that this modification positively affects chromatin
remodeling and thus transcriptional activation. CARM-1 has also
been shown to play a co-activator role in gene transcription
mediated by the nuclear hormone receptor (NHR) family of
transcription factors. In this context, CARM-1 displays an absolute
requirement for the presence a member of the NHR co-activator
family of proteins (SRC-1, GRIP-1 or AIB1) in order to enhance
transcriptional activation by the androgen receptor (AR) or
estrogen receptor (ER). It is also able to co-operate with p300/CBP
and PRMT-1 in NHR dependent transcription. In the resulting
transcriptionally active complex, CARM-1 can methylate both
p300/CBP and the NHR co-activator(s). The fact that mutations of
critical residues in the catalytic domain compromise the
co-activator function of CARM-1 suggests that the integrity of its
methyl transferase domain is not dispensable for its co-activator
function. Additionally, CARM-1 has been implicated to play a key
role in mediating signal transduction pathways among CREB Binding
Protein (CBP), p53 and the muscle enhancer factor-1 (MEF-1).
[0005] In addition to its role as a coactivator in NHR mediated
signaling, CARM-1 has also been implicated as the PRMT responsible
for methylation of PABP (Poly-A-binding protein) and HuR (a member
of the Hu family of protein). PABP and HuR have been shown to bind
to messenger RNAs (e.g. fos, Cox-2, .beta.-catenin) containing the
AUUUUA sequence in the 3' untranslated region thereby stabilizing
the message and leading to increased translation in the
cytoplasm.
[0006] Accordingly, CARM-1 may serve as an oncogene because its
ability to regulate transcription, modulate chromatin organization
and increase the half-life of specific mRNAs. Upon comparison of a
number of matched tumor and normal tissues (lung, colon and
breast), we found that CARM-1 was over-expressed in the tumor
specimens compared to normal tissue (WO03102143). Moreover, Hong et
al (Cancer, 101 (1), 83-89 (2004)) reports that CARM-1 levels are
significantly higher in PIN (Prostatic intraepithelial neoplasia)
and prostatic adenocarcinoma specimens from patients compared to
benign prostate tissue specimens. See "Aberrant expression of
CARM-1, a transcriptional coactivator of Androgen receptor in the
development of prostate carcinoma and androgen independent status.
In twelve patients with androgen independent prostatic
adenocarcinoma, the expression of CARM-1 was significantly
increased when compared to patients without previous hormonal
treatment.
[0007] Furthermore, HuR, one of the substrates of CARM-1, has also
been implicated in cancer (Li et al, J Biol Chem, 277, 44623
(2002), Erkinheimo et al, Cancer Res, 63, 7591 (2003)) HuR is a
nuclear protein but is predominantly cytoplasmic in tumor cells.
Increased cytoplasmic presence predicts a poor prognosis. Thus,
methylation by CARM-1 may be responsible for the cytoplasmic
presence of HuR which resulted in the increased stabilization of
mRNAs of genes implicated in cancer (e.g. fos, Cox-2,
.beta.-catenin) (Denkert et al, Cancer Res, 64, 189 (2004).
Accordingly, it is an attractive therapeutic option for cancer
patients to inhibit the enzymatic function of CARM-1 by using a
small organic molecule.
SUMMARY OF THE INVENTION
[0008] The present invention provides a compound of the following
formula I, or a stereoisomer, a tautomer, a pharmaceutically
acceptable salt or solvate thereof, which compounds are especially
useful as inhibitors of PRMTs and/or CARM-1; ##STR2##
[0009] wherein: [0010] Ring Q is phenyl; 5-membered heteroaryl, in
which Z.sub.4 is a bond, Z.sub.1, Z.sub.2, Z.sub.3 and Z.sub.5 are
each independently C, N, O, or S, and at least one of Z.sub.1,
Z.sub.2, Z.sub.3 and Z.sub.5 is a heteroatom selected from N, O and
S; or 6-membered heteroaryl, in which Z.sub.1, Z.sub.2, Z.sub.3,
Z.sub.4 and Z.sub.5 are each independently C, or N, and at least
one of Z.sub.1, Z.sub.2, Z.sub.3, Z.sub.4 and Z.sub.5 is N; [0011]
W is --C(.dbd.O)NR.sub.8--, --NR.sub.9C(.dbd.O)--,
--NR.sub.9C(.dbd.O)NR.sub.8--, ##STR3## [0012] alternatively,
W--(CH.sub.2)--(O)n-R.sub.7 is ##STR4## [0013] R.sub.1 is H,
halogen, CN, alkyl or substituted alkyl, O--C.sub.1-C.sub.4 alkyl,
S--C.sub.1-C.sub.4 alkyl, or SO.sub.2--C.sub.1-C.sub.4 alkyl;
[0014] R.sub.2 is H, or C.sub.1-C.sub.4 alkyl; [0015] R.sub.3 is H,
Me or Et, or optionally R.sub.3 together with R.sub.4 may form a 5-
or 6-membered heterocycle [0016] R.sub.4 is H, Me, Et, iso-propyl,
CH.sub.2Ph, OH, or OPh, or optionally R.sub.4 together with R.sub.3
may form a 5- or 6-membered heterocycle; [0017] R.sub.5 is nil, H,
Me, Et, propyl, iso-propyl, OMe, OEt, SMe, SO.sub.2Me, CF.sub.3, or
OCF.sub.3; [0018] R.sub.6 is nil, H, Me, or Et, or optionally
R.sub.6 together with R.sub.8 may form a 5- or 6-membered
heterocycle; [0019] R.sub.7 is cycloalkyl or substituted
cycloalkyl, heterocycle or substituted heterocycle, or aryl or
substituted aryl; [0020] R.sub.8 is H, or Me, or optionally R.sub.8
together with R.sub.6 may form a 5- or 6-membered heterocycle;
[0021] or alternatively R.sub.8 together with R.sub.7 may form a 5-
or 6-membered heterocycle or substituted heterocycle; [0022]
R.sub.9 is H, or Me; [0023] m is 0, 1, 2 or 3; [0024] n is 0 or 1;
and [0025] p is 1, 2 or 3.
[0026] The present invention further provides a compound of the
following formula I, or a stereoisomer, a tautomer, a
pharmaceutically acceptable salt or solvate thereof, which
compounds are especially useful as inhibitors of PRMTs and/or
CARM-1. ##STR5##
[0027] As used in formula I, and throughout the specification, the
symbols have the following meanings unless otherwise indicated, and
are, for each occurrence, independently selected: [0028] Ring Q is
phenyl; 5-membered heteroaryl, in which Z.sub.4 is a bond, Z.sub.1,
Z.sub.2, Z.sub.3 and Z.sub.5 are each independently C, N, O, or S,
and at least one of Z.sub.1, Z.sub.2, Z.sub.3 and Z.sub.5 is a
heteroatom selected from N, O and S; or 6-membered heteroaryl, in
which Z.sub.1, Z.sub.2, Z.sub.3, Z.sub.4 and Z.sub.5 are each
independently C, or N, and at least one of Z.sub.1, Z.sub.2,
Z.sub.3, Z.sub.4 and Z.sub.5 is N; [0029] W is
--C(.dbd.O)NR.sub.8--, --NR.sub.9C(.dbd.O)--, ##STR6## [0030]
R.sub.1 is H, halogen, CN, alkyl or substituted alkyl,
O--C.sub.1-C.sub.4 alkyl, S--C.sub.1-C.sub.4 alkyl, or
SO.sub.2--C.sub.1-C.sub.4 alkyl; [0031] R.sub.2 is H or
C.sub.1-C.sub.4 alkyl; [0032] R.sub.3 is H, Me or Et, or optionally
R.sub.3 together with R.sub.4 may form a 5- or 6-membered
heterocycle [0033] R.sub.4 is H, Me, Et, iso-propyl, CH.sub.2Ph,
OH, or OPh, or optionally R.sub.4 together with R.sub.3 may form a
5- or 6-membered heterocycle; [0034] R.sub.5 is nil, H, Me, Et,
propyl, iso-propyl, OMe, OEt, SMe, SO.sub.2Me, CF.sub.3, or
OCF.sub.3; [0035] R.sub.6 is nil, H, Me, or Et, or optionally
R.sub.6 together with R.sub.8 may form a 5- or 6-membered
heterocycle; [0036] R.sub.7 is cycloalkyl or substituted
cycloalkyl, heterocycle or substituted heterocycle, or aryl or
substituted aryl; [0037] R.sub.8 is H, or Me, or optionally R.sub.8
together with R.sub.6 may form a 5- or 6-membered heterocycle;
[0038] or alternatively R.sub.8 together with R.sub.7 may form a 5-
or 6-membered heterocycle or substituted heterocycle; [0039]
R.sub.9 is H, or Me; [0040] m is 0, 1, 2 or 3; [0041] n is 0 or 1;
and [0042] p is 1, 2 or 3.
[0043] The present invention also relates to methods of using
compounds of formula I in the treatment of hyperproliferative,
inflammatory, infectious, and immunoregulatory disorders and
diseases, and to pharmaceutical compositions containing such
compounds.
[0044] The present invention provides pharmaceutical compositions
comprising a pharmaceutically acceptable carrier and a
therapeutically effective amount of at least one of the compounds
of the present invention or a pharmaceutically acceptable salt or
prodrug form thereof.
[0045] The present invention provides pharmaceutical compositions
comprising a pharmaceutically acceptable carrier and an amount of a
compound of formula (I) to provide a therapeutically effective
amount of at least one of the compounds of the present invention or
a pharmaceutically acceptable salt or prodrug form thereof.
[0046] The present invention provides a method of treating cancers,
wherein the cancer is selected from breast cancer, lung cancer,
ovarian cancer, prostate cancer, leukemia, lymphoma, glioblastoma,
brain cancer, melanoma, and colon cancer.
[0047] The present invention provides novel compounds for use in
therapy.
[0048] The present invention provides the use of novel compounds
for the manufacture of a medicament for the treatment of
oncological or immunological diseases.
[0049] In another embodiment, the present invention is directed to
compounds of formula I wherein ring Q is 5-membered heteroaryl, in
which Z.sub.4 is a bond, Z.sub.1, Z.sub.2, Z.sub.3 and Z.sub.5 are
each independently C, N, O, or S, and at least one of Z.sub.1,
Z.sub.2, Z.sub.3 and Z.sub.5 is a heteroatom selected from N, O and
S; or 6-membered heteroaryl, in which Z.sub.1, Z.sub.2, Z.sub.3,
Z.sub.4 and Z.sub.5 are each independently C, or N, and at least
one of Z.sub.1, Z.sub.2, Z.sub.3, Z.sub.4 and Z.sub.5 is N.
[0050] In another embodiment, the present invention is directed to
compounds of formula I wherein ring Q is 5-membered heteroaryl, in
which Z.sub.4 is a bond, Z.sub.1, Z.sub.2, Z.sub.3 and Z.sub.5 are
each independently C, N, O, or S, and at least one of Z.sub.1,
Z.sub.2, Z.sub.3 and Z.sub.5 is a heteroatom selected from N, O and
S.
[0051] In another embodiment, the present invention is directed to
compounds of formula I wherein Z.sub.4 is a bond, Z.sub.1 and
Z.sub.2, are each independently N; Z.sub.3 and Z.sub.5 are each
independently C.
[0052] In another embodiment, the present invention is directed to
compounds of formula I wherein R.sub.7 is aryl or substituted
aryl.
[0053] In another embodiment, the present invention is directed to
compounds of formula I wherein R.sub.7 is phenyl or substituted
phenyl.
[0054] In another embodiment, the present invention is directed to
compounds of formula I wherein W is C(.dbd.O)NR.sub.8--.
[0055] In another embodiment, the present invention is directed to
compounds of formula I wherein W is ##STR7##
[0056] In another embodiment, the present invention is directed to
compounds of formula I wherein W is ##STR8##
[0057] In another embodiment, the present invention is directed to
compounds of formula I wherein R.sub.2 is H, R.sub.3 is H, R.sub.4
is Me, Et, OH, or Ph.
[0058] In another embodiment, the present invention is directed to
compounds of formula I wherein R.sub.4 is Me.
[0059] In another embodiment, the present invention is directed to
compounds of formula I wherein R.sub.4 is (S--) Me.
[0060] In another embodiment, the present invention is directed to
compounds of formula I wherein W is --C(.dbd.O)NR.sub.8--.
[0061] In another embodiment, the present invention is directed to
compounds of formula I wherein Q is ##STR9##
[0062] In another embodiment, the present invention is directed to
compounds of formula I wherein Q is ##STR10##
[0063] In another embodiment, the present invention is directed to
compounds of formula I wherein Q is ##STR11##
[0064] In another embodiment, the present invention is directed to
compounds of formula I wherein Q is ##STR12##
[0065] In another embodiment, the present invention is directed to
compounds of formula I wherein Q is ##STR13##
[0066] In another embodiment, the present invention is directed to
compounds of formula I wherein Q is ##STR14##
[0067] In another embodiment, the present invention is directed to
compounds of formula I wherein Q is ##STR15##
[0068] The invention may be embodied in other specific forms
without departing from the spirit or essential attributes thereof.
This invention also encompasses all combinations of preferred
aspects of the invention noted herein. It is understood that any
and all embodiments of the present invention may be taken in
conjunction with any other embodiment to describe additional even
more preferred embodiments of the present invention. Furthermore,
any elements of an embodiment are meant to be combined with any and
all other elements from any of the embodiments to describe
additional embodiments.
FURTHER DESCRIPTION OF THE INVENTION
Definitions
[0069] The following are definitions of terms used in the present
specification. The initial definition provided for a group or term
herein applies to that group or term throughout the present
specification individually or as part of another group, unless
otherwise indicated.
[0070] The terms "alkyl" and "alk" refers to a straight or branched
chain alkane (hydrocarbon) radical containing from 1 to 12 carbon
atoms, preferably 1 to 6 carbon atoms. Exemplary "alkyl" groups
include methyl, ethyl, propyl, isopropyl, n-butyl, t-butyl,
isobutyl pentyl, hexyl, isohexyl, heptyl, 4,4-dimethylpentyl,
octyl, 2,2,4-trimethylpentyl, nonyl, decyl, undecyl, dodecyl, and
the like. The term "C.sub.1-C.sub.4 alkyl" refers to a straight or
branched chain alkane (hydrocarbon) radical containing from 1 to 4
carbon atoms, such as methyl, ethyl, propyl, isopropyl, n-butyl,
t-butyl, and isobutyl. "Haloalkyl" refers to an alkyl group
substituted with one or more halogen. "Substituted alkyl" refers to
an alkyl group substituted with one or more substituents,
preferably 1 to 4 substituents, at any available point of
attachment. Exemplary substituents include but are not limited to
one or more of the following groups: hydrogen, halogen (e.g., a
single halogen substituent or multiple halo substitutents forming,
in the latter case, groups such as CF.sub.3 or an alkyl group
bearing Cl.sub.3), cyano, nitro, cycloalkyl, alkenyl, cycloalkenyl,
alkynyl, heterocycle, aryl, OR.sub.a, SR.sub.a, S(.dbd.O)R.sub.e,
S(.dbd.O).sub.2R.sub.e, P(.dbd.O).sub.2R.sub.e,
S(.dbd.O).sub.2OR.sub.e, P(.dbd.O).sub.2OR.sub.e, NR.sub.bR.sub.c,
NR.sub.bS(.dbd.O).sub.2R.sub.e, NR.sub.bP(.dbd.O).sub.2R.sub.e,
S(.dbd.O).sub.2NR.sub.bR.sub.c, P(.dbd.O).sub.2NR.sub.bR.sub.c,
C(.dbd.O)OR.sub.e, C(.dbd.O)R.sub.a, C(.dbd.O)NR.sub.bR.sub.c,
OC(.dbd.O)R.sub.a, OC(.dbd.O)NR.sub.bR.sub.c,
NR.sub.bC(.dbd.O)OR.sub.e, NR.sub.dC(.dbd.O)NR.sub.bR.sub.c,
NR.sub.dS(.dbd.O).sub.2NR.sub.bR.sub.c,
NR.sub.dP(.dbd.O).sub.2NR.sub.bR.sub.c, NR.sub.bC(.dbd.O)R.sub.a,
or NR.sub.bP(.dbd.O).sub.2R.sub.e, wherein R.sub.a is hydrogen,
alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, or
aryl; R.sub.b, R.sub.c and R.sub.d are independently hydrogen,
alkyl, cycloalkyl, heterocycle, aryl, or said R.sub.b and R.sub.c
together with the N to which they are bonded optionally form a
heterocycle; and R.sub.e is alkyl, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, heterocycle, or aryl. In the aforementioned
exemplary substitutents, groups such as alkyl, cycloalkyl, alkenyl,
alkynyl, cycloalkenyl, heterocycle and aryl can themselves be
optionally substituted.
[0071] The term "alkenyl" refers to a straight or branched chain
hydrocarbon radical containing from 2 to 12 carbon atoms and at
least one carbon-carbon double bond. Exemplary such groups include
ethenyl or allyl. "Substituted alkenyl" refers to an alkenyl group
substituted with one or more substituents, preferably 1 to 4
substituents, at any available point of attachment. Exemplary
substituents include, but are not limited to, alkyl or substituted
alkyl, as well as those groups recited above as exemplary alkyl
substituents.
[0072] The term "alkynyl" refers to a straight or branched chain
hydrocarbon radical containing from 2 to 12 carbon atoms and at
least one carbon to carbon triple bond. Exemplary such groups
include ethynyl. "Substituted alkynyl" refers to an alkynyl group
substituted with one or more substituents, preferably 1 to 4
substituents, at any available point of attachment. Exemplary
substituents include, but are not limited to, alkyl or substituted
alkyl, as well as those groups recited above as exemplary alkyl
substituents.
[0073] The term "cycloalkyl" refers to a fully saturated cyclic
hydrocarbon group containing from 1 to 4 rings and 3 to 8 carbons
per ring. Exemplary such groups include cyclopropyl, cyclobutyl,
cyclopentyl, cyclohexyl, cycloheptyl, etc. "Substituted cycloalkyl"
refers to a cycloalkyl group substituted with one or more
substituents, preferably 1 to 4 substituents, at any available
point of attachment. Exemplary substituents include, but are not
limited to, nitro, cyano, alkyl or substituted alkyl, as well as
those groups recited above as exemplary alkyl substituents.
Exemplary substituents also include spiro-attached or fused cyclic
substituents, especially spiro-attached cycloalkyl, spiro-attached
cycloalkenyl, spiro-attached heterocycle (excluding heteroaryl),
fused cycloalkyl, fused cycloalkenyl, fused heterocycle, or fused
aryl, where the aforementioned cycloalkyl, cycloalkenyl,
heterocycle and aryl substitutents can themselves be optionally
substituted.
[0074] The term "cycloalkenyl" refers to a partially unsaturated
cyclic hydrocarbon group containing 1 to 4 rings and 3 to 8 carbons
per ring. Exemplary such groups include cyclobutenyl,
cyclopentenyl, cyclohexenyl, etc. "Substituted cycloalkenyl" refers
to a cycloalkenyl group substituted with one more substituents,
preferably 1 to 4 substituents, at any available point of
attachment. Exemplary substituents include but are not limited to
nitro, cyano, alkyl or substituted alkyl, as well as those groups
recited above as exemplary alkyl substituents. Exemplary
substituents also include spiro-attached or fused cyclic
substituents, especially spiro-attached cycloalkyl, spiro-attached
cycloalkenyl, spiro-attached heterocycle (excluding heteroaryl),
fused cycloalkyl, fused cycloalkenyl, fused heterocycle, or fused
aryl, where the aforementioned cycloalkyl, cycloalkenyl,
heterocycle and aryl substituents can themselves be optionally
substituted.
[0075] The term "aryl" refers to cyclic, aromatic hydrocarbon
groups that have 1 to 5 aromatic rings, especially monocyclic or
bicyclic groups such as phenyl, biphenyl or naphthyl. Where
containing two or more aromatic rings (bicyclic, etc.), the
aromatic rings of the aryl group may be joined at a single point
(e.g., biphenyl), or fused (e.g., naphthyl, phenanthrenyl and the
like). "Substituted aryl" refers to an aryl group substituted by
one or more substituents, preferably 1 to 3 substituents, at any
point of attachment. Exemplary substituents include, but are not
limited to, nitro, cycloalkyl or substituted cycloalkyl,
cycloalkenyl or substituted cycloalkenyl, cyano, alkyl or
substituted alkyl, as well as those groups recited above as
exemplary alkyl substituents. Exemplary substituents also include
fused cyclic groups, especially fused cycloalkyl, fused
cycloalkenyl, fused heterocycle, or fused aryl, where the
aforementioned cycloalkyl, cycloalkenyl, heterocycle and aryl
substituents can themselves be optionally substituted. Exemplary
aryl and phenyl substituents include but are not limited to one or
more of the following groups: hydrogen, alkyl, haloalkyl, halogen
(e.g., a single halogen substituent or multiple halo substitutents
forming, in the latter case, groups such as CF.sub.3 or an alkyl
group bearing Cl.sub.3), cyano, nitro, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, heterocycle, aryl, OR.sub.a, SR.sub.a,
S(.dbd.O)R.sub.e, S(.dbd.O).sub.2R.sub.e, P(.dbd.O).sub.2R.sub.e,
S(.dbd.O).sub.2OR.sub.e, P(.dbd.O).sub.2OR.sub.e, NR.sub.bR.sub.c,
NR.sub.bS(.dbd.O).sub.2R.sub.e, NR.sub.bP(.dbd.O).sub.2R.sub.e,
S(.dbd.O).sub.2NR.sub.bR.sub.c, P(.dbd.O).sub.2NR.sub.bR.sub.c,
C(.dbd.O)OR.sub.e, C(.dbd.O)R.sub.a, C(.dbd.O)NR.sub.bR.sub.c,
OC(.dbd.O)R.sub.a, OC(.dbd.O)NR.sub.bR.sub.c,
NR.sub.bC(.dbd.O)OR.sub.e, NR.sub.dC(.dbd.O)NR.sub.bR.sub.c,
NR.sub.dS(.dbd.O).sub.2NR.sub.bR.sub.c,
NR.sub.dP(.dbd.O).sub.2NR.sub.bR.sub.c, NR.sub.bC(.dbd.O)R.sub.a,
or NR.sub.bP(.dbd.O).sub.2R.sub.e, wherein R.sub.a is hydrogen,
alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, or
aryl; R.sub.b, R.sub.c and R.sub.d are independently hydrogen,
alkyl, cycloalkyl, heterocycle, aryl, or said R.sub.b and R.sub.c
together with the N to which they are bonded optionally form a
heterocycle; and R.sub.e is alkyl, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, heterocycle, or aryl. In the aforementioned
exemplary substitutents, groups such as alkyl, cycloalkyl, alkenyl,
alkynyl, cycloalkenyl, heterocycle and aryl can themselves be
optionally substituted.
[0076] The terms "heterocycle" and "heterocyclic" refer to fully
saturated, or partially or fully unsaturated, including aromatic
(i.e., "heteroaryl") cyclic groups (for example, 4 to 7 membered
monocyclic, 7 to 11 membered bicyclic, or 10 to 16 membered
tricyclic ring systems) which have at least one heteroatom in at
least one carbon atom-containing ring. Each ring of the
heterocyclic group containing a heteroatom may have 1, 2, 3, or 4
heteroatoms selected from nitrogen atoms, oxygen atoms and/or
sulfur atoms, where the nitrogen and sulfur heteroatoms may
optionally be oxidized and the nitrogen heteroatoms may optionally
be quaternized. (The term "heteroarylium" refers to a heteroaryl
group bearing a quaternary nitrogen atom and thus a positive
charge.) The heterocyclic group may be attached to the remainder of
the molecule at any heteroatom or carbon atom of the ring or ring
system. Exemplary monocyclic heterocyclic groups include
azetidinyl, pyrrolidinyl, pyrrolyl, pyrazolyl, oxetanyl,
pyrazolinyl, imidazolyl, imidazolinyl, imidazolidinyl, oxazolyl,
oxazolidinyl, isoxazolinyl, isoxazolyl, thiazolyl, thiadiazolyl,
thiazolidinyl, isothiazolyl, isothiazolidinyl, furyl,
tetrahydrofuryl, thienyl, oxadiazolyl, piperidinyl, piperazinyl,
2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolodinyl,
2-oxoazepinyl, azepinyl, hexahydrodiazepinyl, 4-piperidonyl,
pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, triazinyl, triazolyl,
tetrazolyl, tetrahydropyranyl, morpholinyl, thiamorpholinyl,
thiamorpholinyl sulfoxide, thiamorpholinyl sulfone, 1,3-dioxolane
and tetrahydro-1,1-dioxothienyl, and the like. Exemplary bicyclic
heterocyclic groups include indolyl, isoindolyl, benzothiazolyl,
benzoxazolyl, benzoxadiazolyl, benzothienyl, quinuclidinyl,
quinolinyl, tetrahydroisoquinolinyl, isoquinolinyl, benzimidazolyl,
benzopyranyl, indolizinyl, benzofuryl, benzofurazanyl, chromonyl,
coumarinyl, benzopyranyl, cinnolinyl, quinoxalinyl, indazolyl,
pyrrolopyridyl, furopyridinyl (such as furo[2,3-c]pyridinyl,
furo[3,2-b]pyridinyl] or furo[2,3-b]pyridinyl), dihydroisoindolyl,
dihydroquinazolinyl (such as 3,4-dihydro-4-oxo-quinazolinyl),
triazinylazepinyl, tetrahydroquinolinyl and the like. Exemplary
tricyclic heterocyclic groups include carbazolyl, benzidolyl,
phenanthrolinyl, acridinyl, phenanthridinyl, xanthenyl and the
like.
[0077] "Substituted heterocycle" and "substituted heterocyclic"
(such as "substituted heteroaryl") refer to heterocycle or
heterocyclic groups substituted with one or more substituents,
preferably 1 to 4 substituents, at any available point of
attachment. Exemplary substituents include, but are not limited to,
cycloalkyl or substituted cycloalkyl, cycloalkenyl or substituted
cycloalkenyl, nitro, oxo (i.e., .dbd.O), cyano, alkyl or
substituted alkyl, as well as those groups recited above as
exemplary alkyl substituents. Exemplary substituents also include
spiro-attached or fused cyclic substituents at any available point
or points of attachment, especially spiro-attached cycloalkyl,
spiro-attached cycloalkenyl, spiro-attached heterocycle (excluding
heteroaryl), fused cycloalkyl, fused cycloalkenyl, fused
heterocycle, or fused aryl, where the aforementioned cycloalkyl,
cycloalkenyl, heterocycle and aryl substituents can themselves be
optionally substituted. Exemplary substituents include but are not
limited to one or more of the following groups: hydrogen, alkyl,
haloalkyl, halogen (e.g., a single halogen substituent or multiple
halo substitutents forming, in the latter case, groups such as
CF.sub.3 or an alkyl group bearing Cl.sub.3), cyano, nitro,
cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, aryl,
OR.sub.a, SR.sub.a, S(.dbd.O)R.sub.e, S(.dbd.O).sub.2R.sub.e,
P(.dbd.O).sub.2R.sub.e, S(.dbd.O).sub.2OR.sub.e,
P(.dbd.O).sub.2OR.sub.e, NR.sub.bR.sub.c,
NR.sub.bS(.dbd.O).sub.2R.sub.e, NR.sub.bP(.dbd.O).sub.2R.sub.e,
S(.dbd.O).sub.2NR.sub.bR.sub.c, P(.dbd.O).sub.2NR.sub.bR.sub.c,
C(.dbd.O)OR.sub.e, C(.dbd.O)R.sub.a, C(.dbd.O)NR.sub.bR.sub.c,
OC(.dbd.O)R.sub.a, OC(.dbd.O)NR.sub.bR.sub.c,
NR.sub.bC(.dbd.O)OR.sub.e, NR.sub.dC(.dbd.O)NR.sub.bR.sub.c,
NR.sub.dS(.dbd.O).sub.2NR.sub.bR.sub.c,
NR.sub.dP(.dbd.O).sub.2NR.sub.bR.sub.c, NR.sub.bC(.dbd.O)R.sub.a,
or NR.sub.bP(.dbd.O).sub.2R.sub.e, wherein R.sub.a is hydrogen,
alkyl, cycloalkyl, alkenyl, cycloalkenyl, alkynyl, heterocycle, or
aryl; R.sub.b, R.sub.c and R.sub.d are independently hydrogen,
alkyl, cycloalkyl, heterocycle, aryl, or said R.sub.b and R.sub.c
together with the N to which they are bonded optionally form a
heterocycle; and R.sub.e is alkyl, cycloalkyl, alkenyl,
cycloalkenyl, alkynyl, heterocycle, or aryl. In the aforementioned
exemplary substitutents, groups such as alkyl, cycloalkyl, alkenyl,
alkynyl, cycloalkenyl, heterocycle and aryl can themselves be
optionally substituted. The term "quaternary nitrogen" refers to a
tetravalent positively charged nitrogen atom including, for
example, the positively charged nitrogen in a tetraalkylammonium
group (e.g., tetramethylammonium, N-methylpyridinium), the
positively charged nitrogen in protonated ammonium species (e.g.,
trimethyl-hydroammonium, N-hydropyridinium), the positively charged
nitrogen in amine N-oxides (e.g., N-methyl-morpholine-N-oxide,
pyridine-N-oxide), and the positively charged nitrogen in an
N-amino-ammonium group (e.g., N-aminopyridinium).
[0078] The terms "halogen" or "halo" refer to chlorine, bromine,
fluorine or iodine.
[0079] The term "carbocyclic" refers to aromatic or non-aromatic 3
to 7 membered monocyclic and 7 to 11 membered bicyclic groups, in
which all atoms of the ring or rings are carbon atoms. "Substituted
carbocyclic" refers to a carbocyclic group substituted with one or
more substituents, preferably 1 to 4 substituents, at any available
point of attachment. Exemplary substituents include, but are not
limited to, nitro, cyano, OR.sub.a, wherein R.sub.a is as defined
hereinabove, as well as those groups recited above as exemplary
cycloalkyl substituents.
[0080] When a functional group is termed "protected", this means
that the group is in modified form to mitigate, especially
preclude, undesired side reactions at the protected site. Suitable
protecting groups for the methods and compounds described herein
include, without limitation, those described in standard textbooks,
such as Greene, T. W. et al., Protective Groups in Organic
Synthesis, Wiley, N.Y. (1999).
[0081] Unless otherwise indicated, any heteroatom with unsatisfied
valences is assumed to have hydrogen atoms sufficient to satisfy
the valences.
[0082] The compounds of formula I form salts which are also within
the scope of this invention. Reference to a compound of the formula
I herein is understood to include reference to salts thereof,
unless otherwise indicated. The term "salt(s)", as employed herein,
denotes acidic and/or basic salts formed with inorganic and/or
organic acids and bases. In addition, when a compound of formula I
contains both a basic moiety, such as but not limited to a pyridine
or imidazole, and an acidic moiety such as but not limited to a
carboxylic acid, zwitterions ("inner salts") may be formed and are
included within the term "salt(s)" as used herein. Pharmaceutically
acceptable (i.e., non-toxic, physiologically acceptable) salts are
preferred, although other salts are also useful, e.g., in isolation
or purification steps which may be employed during preparation.
Salts of the compounds of the formula I may be formed, for example,
by reacting a compound I with an amount of acid or base, such as an
equivalent amount, in a medium such as one in which the salt
precipitates or in an aqueous medium followed by
lyophilization.
[0083] The compounds of formula I which contain a basic moiety,
such as but not limited to an amine or a pyridine or imidazole
ring, may form salts with a variety of organic and inorganic acids.
Exemplary acid addition salts include acetates (such as those
formed with acetic acid or trihaloacetic acid, for example,
trifluoroacetic acid), adipates, alginates, ascorbates, aspartates,
benzoates, benzenesulfonates, bisulfates, borates, butyrates,
citrates, camphorates, camphorsulfonates, cyclopentanepropionates,
digluconates, dodecylsulfates, ethanesulfonates, fumarates,
glucoheptanoates, glycerophosphates, hemisulfates, heptanoates,
hexanoates, hydrochlorides, hydrobromides, hydroiodides,
hydroxyethanesulfonates (e.g., 2-hydroxyethanesulfonates),
lactates, maleates, methanesulfonates, naphthalenesulfonates (e.g.,
2-naphthalenesulfonates), nicotinates, nitrates, oxalates,
pectinates, persulfates, phenylpropionates (e.g.,
3-phenylpropionates), phosphates, picrates, pivalates, propionates,
salicylates, succinates, sulfates (such as those formed with
sulfuric acid), sulfonates, tartrates, thiocyanates,
toluenesulfonates such as tosylates, undecanoates, and the
like.
[0084] The compounds of formula I which contain an acidic moiety,
such but not limited to a carboxylic acid, may form salts with a
variety of organic and inorganic bases. Exemplary basic salts
include ammonium salts, alkali metal salts such as sodium, lithium
and potassium salts, alkaline earth metal salts such as calcium and
magnesium salts, salts with organic bases (for example, organic
amines) such as benzathines, dicyclohexylamines, hydrabamines
(formed with N,N-bis(dehydroabietyl)ethylenediamine),
N-methyl-D-glucamines, N-methyl-D-glycamides, t-butyl amines, and
salts with amino acids such as arginine, lysine and the like. Basic
nitrogen-containing groups may be quaternized with agents such as
lower alkyl halides (e.g. methyl, ethyl, propyl, and butyl
chlorides, bromides and iodides), dialkyl sulfates (e.g. dimethyl,
diethyl, dibutyl, and diamyl sulfates), long chain halides (e.g.
decyl, lauryl, myristyl and stearyl chlorides, bromides and
iodides), aralkyl halides (e.g. benzyl and phenethyl bromides), and
others.
[0085] Prodrugs and solvates of the compounds of the invention are
also contemplated herein. The term "prodrug" as employed herein
denotes a compound that, upon administration to a subject,
undergoes chemical conversion by metabolic or chemical processes to
yield a compound of the formula I, or a salt and/or solvate
thereof. Solvates of the compounds of formula I include, for
example, hydrates.
[0086] Compounds of the formula I, and salts thereof, may exist in
their tautomeric form (for example, as an amide or imino ether).
All such tautomeric forms are contemplated herein as part of the
present invention.
[0087] All stereoisomers of the present compounds (for example,
those which may exist due to asymmetric carbons on various
substituents), including enantiomeric forms and diastereomeric
forms, are contemplated within the scope of this invention.
Individual stereoisomers of the compounds of the invention may, for
example, be substantially free of other isomers (e.g., as a pure or
substantially pure optical isomer having a specified activity), or
may be admixed, for example, as racemates or with all other, or
other selected, stereoisomers. The chiral centers of the present
invention may have the S or R configuration as defined by the IUPAC
1974 Recommendations. The racemic forms can be resolved by physical
methods, such as, for example, fractional crystallization,
separation or crystallization of diastereomeric derivatives or
separation by chiral column chromatography. The individual optical
isomers can be obtained from the racemates by any suitable method,
including without limitation, conventional methods, such as, for
example, salt formation with an optically active acid followed by
crystallization.
[0088] Compounds of the formula I are, subsequent to their
preparation, may be isolated and purified to obtain a composition
containing an amount by weight equal to or greater than 99% formula
I compound ("substantially pure" compound I), which is then used or
formulated as described herein. Such "substantially pure" compounds
of the formula I are also contemplated herein as part of the
present invention.
[0089] All configurational isomers of the compounds of the present
invention are contemplated, either in admixture or in pure or
substantially pure form. The definition of compounds of the present
invention embraces both cis (Z) and trans (E) alkene isomers, as
well as cis and trans isomers of cyclic hydrocarbon or heterocyclic
rings.
[0090] "Therapeutically effective amount" is intended to include an
amount of a compound of the present invention alone or an amount of
the combination of compounds claimed or an amount of a compound of
the present invention in combination with other active ingredients
effective to inhibit CARM-1 or effective to treat or prevent
inflammatory disorders.
[0091] As used herein, "treating" or "treatment" cover the
treatment of a disease-state in a mammal, particularly in a human,
and include: (a) preventing the disease-state from occurring in a
mammal, in particular, when such mammal is predisposed to the
disease-state but has not yet been diagnosed as having it; (b)
inhibiting the disease-state, i.e., arresting it development;
and/or (c) relieving the disease-state, i.e., causing regression of
the disease state.
[0092] Throughout the specifications, groups and substituents
thereof may be chosen to provide stable moieties and compounds.
Biological Assay
[0093] The pharmacological properties of the compounds of this
invention may be confirmed by a number of pharmacological assays.
The exemplified pharmacological assays which follow have been
carried out with the compounds according to the invention and/or
their pharmaceutically acceptable salts.
[0094] A methylation based filter assay was devised to test
compounds that specifically inhibited CARM 1 dependent methylation.
Human full length CARM1 purified from baculovirus infected insect
cells was used as the source for enzyme. Histone H3 (Roche Applied
Science) was used as the preferred substrate for the assay since
the methylation of several amino acids of Histone H3 by CARM1 has
been well documented. Tritiated S-Adenosyl-L-Methionine (SAM)
(Amersham Pharmacia Biotech) was used as a cofactor since CARM1
exhibits an absolute requirement for SAM for its catalytic
activity. Methylation reactions were performed for 75-90 minutes at
room temperature using enzyme (CARM-1), substrate (Histone H3) and
cofactor (SAM) in the presence of methylation buffer (20 mM
Tris.HCl pH 8.0, 200 mM NaCl, 0.4 mM EDTA) and in the presence or
absence of compound. Reactions were terminated using TCA and
precipitated with BSA overnight. They were harvested the next
morning by passing the reactions through a filter and the filters
washed. The signal on the filters was read in a Top Count after
addition of MicroScint-20.
Protocol for Methylation Based Filter Assay
I. Reaction Mixtures and Solutions:
[0095] 10.times. Methyl transferase Buffer (MTB):
[0096] 20 mM Tris HCl, pH 8.0
[0097] 200 mM NaCl
[0098] 0.4 mM EDTA [0099] 3.times. Test Compound Solution (15 .mu.l
per well): [0100] Test compounds are diluted to 3 fold of final
concentration with 1.times.MTB. For example, if the final
concentration of test compound is designed as 30 .mu.M, then 90
.mu.M compound solution should be made at this step. [0101] Diluted
DMSO Solution (15 .mu.l per well): [0102] 100% DMSO was diluted
with 1.times.MTB to the same concentration as the compound
solution. For example, if the 3.times. concentrate test compound
solution contains 3% DMSO then 3% DMSO should be used for control
and blank wells. [0103] CARM1 Enzyme Mixture (15 .mu.l per well):
[0104] Pre-determined amount of CARM1 Enzyme [0105] 0.03 .mu.l 1M
DTT [0106] 0.03 .mu.l 100 mg/ml BSA [0107] 1.5 .mu.l 10.times.MTB
[0108] 13.39 .mu.l H.sub.2O
[0109] Histone H3/[.sup.3H] SAM Mixture (15 .mu.l per well): [0110]
0.8 .mu.l 1 mg/ml Histone H3 [0111] 0.2 .mu.l 1 mCi/ml [3H] SAM
(65-80 Ci/mmol) [0112] 1.5 .mu.l 10.times.MTB [0113] 12.5 .mu.l
H.sub.2O [0114] Stop Solution (45 .mu.l per well): [0115] 20% TCA
[0116] 100 mM Sodium pyrophosphate II. Reaction Steps: [0117] A
plate layout, which contains blank (no enzyme), positive control
(no compound) and test compound wells was created [0118] The
following reaction mixtures and solutions were added to wells
taking care to avoid cross contamination: [0119] 15 .mu.l
1.times.MTB to each of blank wells [0120] 15 .mu.l diluted DMSO
solution to each of blank and positive control wells [0121] 15
.mu.l 3.times. test compound solution to each of test compound
wells [0122] 15 .mu.l CARM1 Mixture to each of compound and
positive control wells [0123] 15 .mu.l Histone H3/[3H] SAM mixture
to all wells [0124] Plates were incubated for 60 minutes at room
temperature (22-24.degree. C.) [0125] 45 .mu.l of stop solution was
added to all wells [0126] Plates were left overnight at 4.degree.
C. [0127] The next day reaction mixes were harvested to filter
plate with a Unifilter harvester [0128] The filter plates were
washed twice with 10% TCA and 4 times with H.sub.2O [0129] Air dry
filter plate [0130] 30 .mu.l Microscint-20 was added to each well
and plates were covered with Top-seal [0131] Plates were read in
the Top Count and the data was analyzed to generate the IC.sub.50
values. III. The Final Concentration of Components (in 60 .mu.l
total reaction volume): [0132] CARM1 Enzyme=as pre-determined
[0133] Histone H3=0.8 .mu.g (1.16 .mu.M) [0134] [3H] SAM=0.2 .mu.Ci
(56 nM) [0135] DMSO=1%
[0136] Compounds of the present invention have activity in the
above described assay. Compounds of the present invention described
herein have an IC.sub.50 of less than 10 .mu.M.
Protocol for 3H Thymidine Incorporation Assay for IC.sub.50
Evaluation
[0137] Inhibition of tumor cell proliferation upon treatment with
compounds was monitored using the 3H thymidine incorporation assay.
Cells of appropriate density were plated in 96-well plates and
compounds added on the same day. Compound treatment was continued
for either 3 or 6 days and dose response curves were determined as
described below.
[0138] Materials and Reagents RPMI media [0139] Fetal Bovine Serum
[0140] Trypsin-EDTA [0141] DMSO [0142] 96 Deep-well plate, 24/box
[0143] [6-3H]Thymidine, 5 mCi [0144] Unifilter-96, GF-B [0145]
Microscient PS [0146] TopSeal-A:96-Well Microplates [0147] Assay
Media: [0148] RPMI media+2.5% FBS [0149] Compound dilution:
[0150] A 96 deep-well plate was used for compound dilution.
Compounds were diluted 1:3 serially. A 10-point dilution was
performed starting at a concentration of 40 .mu.M (2.times.). Assay
media in columns 2-10 contained 0.4% DMSO (2.times.).
[0151] An example is shown below: TABLE-US-00001 1 2 3 4 5 6 7 8 9
10 Conc. 40 13.333 4.444 1.482 0.494 0.165 0.055 0.018 0.06 0.02
.mu.M(2X) Assay 0.9 0.6 0.6 0.6 0.6 0.6 0.6 0.6 0.6 0.6 media (ml)
10 .mu.M 3.6 Compd (.mu.l) Mix well and sequential dilution
Transfer(ml): 0.3 0.3 0.3 0.3 0.3 0.3 0.3 0.3 0.3
[0152] Preparation of cell suspension solution:
[0153] 2.times.103-4.times.104 cells/ml (depending on cell type) of
cell suspension was prepared in the assay media. Following is an
example: TABLE-US-00002 Day 3 Day 6 Cell line Cells/well Cells/ml
Cells/well Cells/ml A549 500 1.0 .times. 10.sup.4 100 2 .times.
10.sup.3 600 1.2 .times. 10.sup.4 200 4 .times. 10.sup.3 1000 2
.times. 10.sup.4 500 1 .times. 10.sup.4 2000 4 .times. 10.sup.4
1000 2 .times. 10.sup.4
[0154] Plate cells: [0155] 4 compounds were tested per plate.
[0156] 50 .mu.l of 2.times. compounds and 50 .mu.l of cell
suspension were added per well of a 96-well plate. TABLE-US-00003 1
2 3 4 5 6 7 8 9 10 11 12 Conc. 20 6.667 2.222 0.74 0.245 0.082
0.027 0.009 0.003 0.001 0 .mu.M A Compound 1 B C Compound 2 D E
Compound 3 F G Compound 4 H Note: Column 11 was the control. 50 ul
of assay media containing 0.4% DMSO and 50 .mu.l of cell suspension
were added per well.
[0157] On day 3 and day 6: [0158] Addition of [.sup.3H] thymidine
media: [0159] Dilution of [.sup.3H] thymidine solution: 44 .mu.l of
thymidine stock liquid was mixed with 956 .mu.l of assay media for
a total of 1 ml. 10 .mu.l of diluted thymidine solution was added
into each well (final concentration of thymidine is 4 .mu.Ci/well).
[0160] Harvest cells: [0161] Media from the plates was discarded
and 75 .mu.l of 0.25% trypsin-EDTA was added to each well. Plates
were incubated at 37.degree. C. for 15 min. Cells were then
harvested with Unifilter-96 GF-B plate by using the harvester.
[0162] Air dry plate: Plates were dried overnight.
[0163] Data analysis: [0164] The bottom of the plate was sealed
with sealing paper. [0165] 50 .mu.l of Microscint.TM. PS was added
to each well and the top of the plate was sealed with topseal.
[0166] The plate was read on a Topcon instrument and the data was
analyzed to generate the IC.sub.50 value.
[0167] The same protocol was used for the other cancer cell
lines.
[0168] The compounds of the present invention have been tested in
the following cell lines MDA-231 (breast); MDA-453 (breast);
MDA-468 (breast); HS-578T (breast); DU-4475 (breast); BT-549
(breast); MCF-7 (breast;) K562 (leukemia); MolT4 (leukemia;)
CCRF-CEM (leukemia); OCZ-CY19 (lymphoma); SK-Mel5 (melanoma);
SK-Mel28 (melanoma); A549 (lung;) LX1 (lung); H23 (lung); H226
(lung); H522 (lung); H661 (lung); A375 (lung); and MSTO-211H
(lung). Additionally, compounds of the present invention may be
tested in the following cell lines: SW480 (colon); HCT116 (colon);
PC3 (prostate); and LnCaP (prostate).
[0169] Compounds of the present invention have activity in the
above described assay. Compounds of the present invention described
herein have an IC.sub.50 of less than 10 .mu.M.
[0170] In another embodiment of the invention, a method is provided
for treating a proliferative disease via modulation of CARM-1
(PRMT-4) by administering to a patient in need of such treatment an
effective amount of a compound of formula I, as defined above, in
combination (simultaneously or sequentially) with at least one
other anti-cancer agent. In a preferred embodiment, the
proliferative disease is cancer.
[0171] The invention further provides pharmaceutical compositions
comprising compounds having formula I together with a
pharmaceutically acceptable carrier.
[0172] More specifically, the compounds of Formulas I are useful in
the treatment of a variety of cancers, including, but not limited
to, the following:
[0173] a) carcinoma, including that of the bladder, breast, colon,
kidney, liver, lung, including small cell lung cancer, esophagus,
gall bladder, ovary, pancreas, stomach, cervix, thyroid, prostate,
and skin, including squamous cell carcinoma;
[0174] b) hematopoietic tumors of lymphoid lineage, including
leukemia, acute lymphocytic leukemia, acute lymphoblastic leukemia,
B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkins
lymphoma, hairy cell lymphoma and Burkett's lymphoma;
[0175] c) hematopoietic tumors of myeloid lineage, including acute
and chronic myelogenous leukemias, myelodysplastic syndrome and
promyelocytic leukemia;
[0176] d) tumors of mesenchymal origin, including fibrosarcoma and
rhabdomyosarcoma;
[0177] e) tumors of the central and peripheral nervous system,
including astrocytoma, neuroblastoma, glioma and schwannomas;
and
[0178] f) other tumors, including melanoma, seminoma,
teratocarcinoma, osteosarcoma, xenoderoma pigmentosum,
keratoctanthoma, thyroid follicular cancer and Kaposi's
sarcoma.
[0179] The compounds of Formula I are useful in the treatment of
breast cancer, leukemia, melanoma, lung cancer, colon cancer and
prostate cancer.
[0180] The compounds of the present invention may be employed alone
or in combination with each other and/or other suitable therapeutic
agents useful in the treatment of cancer such as PTK inhibitors,
antiinflammatories, antiproliferatives, chemotherapeutic agents,
immunosuppressants, anticancer agents and cytotoxic agents.
[0181] Exemplary classes of anti-cancer agents and cytotoxic agents
include, but are not limited to: alkylating agents, such as
nitrogen mustards, alkyl sulfonates, nitrosoureas, ethylenimines,
and triazenes; antimetabolites, such as folate antagonists, purine
analogues, and pyrimidine analogues; antibiotics, such as
anthracyclines (e.g., daunorubicin, doxorubicin), cytarabine
(ara-C; Cytosar-U.RTM.); 6-thioguanine (Tabloid.RTM.), mitoxantrone
(Novantrone.RTM.) and etoposide (VePesid.RTM.), amsacrine (AMSA),
and all-trans retinoic acid (ATRA), bleomycins, mitomycin,
dactinomycin, and plicamycin; enzymes, such as L-asparaginase;
farnesyl-protein transferase inhibitors; hormonal agents, such as
glucocorticoids, estrogens/antiestrogens, androgens/antiandrogens,
progestins, and luteinizing hormone-releasing hormone anatagonists,
octreotide acetate; microtubule-disruptor agents, such as
ecteinascidins or their analogs and derivatives;
microtubule-stabilizing agents such as paclitaxel (Taxol.RTM.),
docetaxel (Taxotere.RTM.), and epothilones A-F or their analogs or
derivatives; plant-derived products, such as vinca alkaloids,
epipodophyllotoxins, taxanes; and topoisomerase inhibitors;
prenyl-protein transferase inhibitors; and miscellaneous agents
such as, hydroxyurea, procarbazine, mitotane, hexamethylmelamine,
platinum coordination complexes such as cisplatin and carboplatin;
and other agents used as anti-cancer and cytotoxic agents such as
biological response modifiers, growth factors; immune modulators,
and monoclonal antibodies. The compounds of the invention may also
be used in conjunction with radiation therapy.
[0182] Representative examples of these classes of anti-cancer and
cytotoxic agents include, but are not limited to, mechlorethamine
hydrochloride, cyclophosphamide, chlorambucil, melphalan,
ifosfamide, busulfan, carmustin, lomustine, semustine,
streptozocin, thiotepa, dacarbazine, methotrexate, thioguanine,
mercaptopurine, fludarabine, pentastatin, cladribin, cytarabine,
fluorouracil, doxorubicin hydrochloride, daunorubicin, idarubicin,
bleomycin sulfate, mitomycin C, actinomycin D, safracins,
saframycins, quinocarcins, discodermolides, vincristine,
vinblastine, vinorelbine tartrate, etoposide, teniposide,
paclitaxel, tamoxifen, estramustine, estramustine phosphate sodium,
flutamide, buserelin, leuprolide, pteridines, diyneses, levamisole,
aflacon, interferon, interleukins, aldesleukin, filgrastim,
sargramostim, rituximab, BCG, tretinoin, irinotecan hydrochloride,
betamethosone, gemcitabine hydrochloride, altretamine, and topoteca
and any analogs or derivatives thereof.
[0183] Other members of these classes include, but are not limited
to paclitaxel, cisplatin, carboplatin, doxorubicin, carminomycin,
daunorubicin, aminopterin, methotrexate, methopterin, mitomycin C,
ecteinascidin 743, porfiromycin, 5-fluorouracil, 6-mercaptopurine,
gemcitabine, cytosine arabinoside, podophyllotoxin or
podophyllotoxin derivatives such as etoposide, etoposide phosphate
or teniposide, melphalan, vinblastine, vincristine, leurosidine,
vindesine, and leurosine.
[0184] In a combination, the other component(s) of such conjoint
treatment in addition to the antiproliferative treatment defined
herein before may be: surgery, radiotherapy or chemotherapy. Such
chemotherapy may cover three main categories of therapeutic agent:
[0185] (i) antiangiogenic agents that work by different mechanisms
from those defined hereinbefore (for example, linomide, inhibitors
of integrin .alpha.v.beta.3 function, angiostatin, razoxane);
[0186] (ii) cytostatic agents such as antiestrogens (for example,
tamoxifen, toremifene, raloxifene, droloxifene, iodoxifene),
progestogens (for example megestrol acetate), aromatase inhibitors
(for example, anastrozole, letrozole, borazole, exemestane),
antihormones, antiprogestogens, antiandrogens (for example,
flutamide, nilutamide, bicalutamide, cyproterone acetate), LHRH
agonists and antagonists (for example, gosereline acetate,
leuprolide), inhibitors of testosterone 5.alpha.-dihydroreductase
(for example, finasteride), farnesyltransferase inhibitors,
anti-invasion agents (for example, metalloproteinase inhibitors
like marimastat and inhibitors of urokinase plasminogen activator
receptor function) and inhibitors of growth factor function, (such
growth factors include for example, EGF, FGF, platelet derived
growth factor and hepatocyte growth factor such inhibitors include
growth factor antibodies, growth factor receptor antibodies such as
Avastin.RTM. (bevacizumab) and Erbitux.RTM. (cetuximab); tyrosine
kinase inhibitors and serine/threonine kinase inhibitors such as
Gleevec.RTM. (imatinib mesylate)); and [0187] (iii)
antiproliferative/antineoplastic drugs and combinations thereof, as
used in medical oncology, such as antimetabolites (for example,
antifolates like methotrexate, fluoropyrimidines like
5-fluorouracil, purine and adenosine analogues, cytosine
arabinoside); Intercalating antitumour antibiotics (for example,
anthracyclines like doxorubicin, daunomycin, epirubicin and
idarubicin, mitomycin-C, dactinomycin, mithramycin); platinum
derivatives (for example, cisplatin, carboplatin); alkylating
agents (for example, nitrogen mustard, melphalan, chlorambucil,
busulphan, cyclophosphamide, ifosfamide nitrosoureas, thiotepa;
antimitotic agents (for example, vinca alkaloids like vincristine,
vinblastine and vinflunine, and taxoids like Taxol.RTM.
(paclitaxel), Taxotere.RTM. (docetaxel) and newer microbtubule
agents such as epothilone analogs, discodermolide analogs, and
eleutherobin analogs); topoisomerase inhibitors (for example,
epipodophyllotoxins like etoposide and teniposide, amsacrine,
topotecan, CPT-11); cell cycle inhibitors (for example,
flavopyridols); biological response modifiers and proteasome
inhibitors such as Velcade.RTM. (bortezomib).
[0188] The term "anticancer" agent includes any known agent that is
useful for the treatment of cancer including the following:
17.alpha.-ethinylestradiol, diethylstilbestrol, testosterone,
prednisone, fluoxymesterone, dromostanolone propionate,
testolactone, megestrolacetate, methylprednisolone,
methyl-testosterone, prednisolone, triamcinolone, chlorotrianisene,
hydroxyprogesterone, aminoglutethimide, estramustine,
medroxyprogesteroneacetate, leuprolide, flutamide, toremifene,
Zoladex; matrix metalloproteinase inhibitors; VEGF inhibitors, such
as anti-VEGF antibodies (Avastin) and small molecules such as
ZD6474, AZD-2171, SU6668; Vatalanib, BAY-43-9006, SU11248,
CP-547632, and CEP-7055; Her 1 and Her 2 inhibitors including
anti-Her2 antibodies (Herceptin); EGFR inhibitors including
gefitinib, erlotinib, ABX-EGF, EMD72000, 11F8, and cetuximab; Eg5
inhibitors, such as SB-715992, SB-743921, and MKI-833; pan Her
inhibitors, such as canertinib, EKB-569, CI-1033, AEE-788, XL-647,
mAb 2C4, and GW-572016; Src kinase inhibitors such as BMS-354825,
AZD-0530, SKI-606, and AP-23464; Bcr-Abl inhibitors such as
imatinib and AMN107; Casodex.RTM. (bicalutamide, Astra Zeneca),
Tamoxifen; MEK-1 kinase inhibitors, MAPK kinase inhibitors, PI3
kinase inhibitors; Met inhibitors, Aurora kinase inhibitors, PDGF
inhibitors; anti-angiogenic and antivascular agents which, by
interrupting blood flow to solid tumors, render cancer cells
quiescent by depriving them of nutrition; castration, which renders
androgen dependent carcinomas non-proliferative; IGF1R inhibitors
such as those disclosed in US2004/44203A1, inhibitors of
non-receptor and receptor tyrosine kinases; inhibitors of integrin
signaling; tubulin acting agents such as vinblastine, vincristine,
vinorelbine, vinflunine, paclitaxel, docetaxel,
7-O-methylthiomethylpaclitaxel,
4-desacetyl-4-methylcarbonatepaclitaxel,
3'-tert-butyl-3'-N-tert-butyloxycarbonyl-4-deacetyl-3'-dephenyl-3'-N-debe-
nzoyl-4-O-methoxycarbonyl-paclitaxel, C-4 methyl carbonate
paclitaxel, epothilone A, epothilone B, epothilone C, epothilone D,
desoxyepothilone A, desoxyepothilone B, ixabepilone,
[1S-[1R*,3R*(E),7R*,10S*,11R*,12R*,16S*]]-3-[2-[2-(aminomethyl)-4-thiazol-
yl]-1-methylethenyl]-7,11-dihydroxy-8,8,10,12,16-pentamethyl-4-17-dioxabic-
yclo[14.1.0]-heptadecane-5,9-dione, and derivatives thereof; CDK
inhibitors, antiproliferative cell cycle inhibitors,
epidophyllotoxin, etoposide, VM-26; antineoplastic enzymes, e.g.,
topoisomerase I inhibitors, camptothecin, topotecan, SN-38;
procarbazine; mitoxantrone; platinum coordination complexes such as
cisplatin, carboplatin and oxaliplatin; biological response
modifiers; growth inhibitors; antihormonal therapeutic agents;
leucovorin; tegafur; antimetabolites such as purine antagonists
(e.g. 6-thioguanine and 6-mercaptopurine; glutamine antagonists,
e.g. DON (AT-125; d-oxo-norleucine); ribonucleotide reductase
inhibitors; mTOR inhibitors; and haematopoietic growth factors.
[0189] Additional cytotoxic agents include, cyclophosphamide,
doxorubicin, daunorubicin, mitoxanthrone, melphalan, hexamethyl
melamine, thiotepa, cytarabin, idatrexate, trimetrexate,
dacarbazine, L-asparaginase, bicalutamide, leuprolide,
pyridobenzoindole derivatives, interferons, and interleukins.
[0190] In cases where it is desirable to render aberrantly
proliferative cells quiescent in conjunction with or prior to
treatment with the chemotherapeutic methods of the invention,
hormones and steroids (including synthetic analogs):
17a-Ethinylestradiol, Diethylstilbestrol, Testosterone, Prednisone,
Fluoxymesterone, Dromostanolone propionate, Testolactone,
Megestrolacetate, Methylprednisolone, Methyl-testosterone,
Prednisolone, Triamcinolone, hlorotrianisene, Hydroxyprogesterone,
Aminoglutethimide, Estramustine, Medroxyprogesteroneacetate,
Leuprolide, Flutamide, Toremifene, Zoladex can also be administered
to the patient.
[0191] Also suitable for use in the combination chemotherapeutic
methods of the invention are antiangiogenics such as matrix
metalloproteinase inhibitors, and other VEGF inhibitors, such as
anti-VEGF antibodies and small molecules such as ZD6474 and SU6668
are also included. Anti-Her2 antibodies from Genetech may also be
utilized. A suitable EGFR inhibitor is EKB-569 (an irreversible
inhibitor). Also included are Imclone antibody C225 immunospecific
for the EGFR, and src inhibitors.
[0192] Also suitable for use as an antiproliferative cytostatic
agent is Casodex.TM. which renders androgen-dependent carcinomas
non-proliferative. Yet another example of a cytostatic agent is the
antiestrogen Tamoxifen which inhibits the proliferation or growth
of estrogen dependent breast cancer. Inhibitors of the transduction
of cellular proliferative signals are cytostatic agents. Examples
are epidermal growth factor inhibitors, Her-2 inhibitors, MEK-1
kinase inhibitors, MAPK kinase inhibitors, PI3 inhibitors, Src
kinase inhibitors, and PDGF inhibitors.
[0193] As mentioned, certain anti-proliferative agents are
anti-angiogenic and antivascular agents and, by interrupting blood
flow to solid tumors, render cancer cells quiescent by depriving
them of nutrition. Castration, which also renders androgen
dependent carcinomas non-proliferative, may also be utilized.
Starvation by means other than surgical disruption of blood flow is
another example of a cytostatic agent. A particular class of
antivascular cytostatic agents is the combretastatins. Other
exemplary cytostatic agents include MET kinase inhibitors, MAP
kinase inhibitors, inhibitors of non-receptor and receptor tyrosine
kinases, inhibitors of integrin signaling, and inhibitors of
insulin-like growth factor receptors.
[0194] Compounds of Formulas I as modulators of apoptosis, will be
useful in the treatment of cancer (including but not limited to
those types mentioned herein above), viral infections (including
but not limited to herpevirus, poxvirus, Epstein-Barr virus,
Sindbis virus and adenovirus), prevention of AIDS development in
HIV-infected individuals, autoimmune diseases (including but not
limited to systemic lupus, erythematosus, autoimmune mediated
glomerulonephritis, rheumatoid arthritis, psoriasis, inflammatory
bowel disease, and autoimmune diabetes mellitus), neurodegenerative
disorders (including but not limited to Alzheimer's disease,
AIDS-related dementia, Parkinson's disease, amyotrophic lateral
sclerosis, retinitis pigmentosa, spinal muscular atrophy and
cerebellar degeneration), myelodysplastic syndromes, aplastic
anemia, ischemic injury associated with myocardial infarctions,
stroke and reperfusion injury, arrhythmia, atherosclerosis,
toxin-induced or alcohol related liver diseases, hematological
diseases (including but not limited to chronic anemia and aplastic
anemia), degenerative diseases of the musculoskeletal system
(including but not limited to osteoporosis and arthritis)
aspirin-sensitive rhinosinusitis, cystic fibrosis, multiple
sclerosis, kidney diseases and cancer pain.
[0195] Compounds of Formulas I may modulate the level of cellular
RNA and DNA synthesis. These agents would therefore be useful in
the treatment of viral infections (including but not limited to
HIV, human papilloma virus, herpesvirus, poxvirus, Epstein-Barr
virus, Sindbis virus and adenovirus).
[0196] Compounds of Formulas I may be useful in the chemoprevention
of cancer. Chemoprevention is defined as inhibiting the development
of invasive cancer by either blocking the initiating mutagenic
event or by blocking the progression of pre-malignant cells that
have already suffered an insult or inhibiting tumor relapse.
[0197] Compounds of Formulas I may also be useful in inhibiting
tumor angiogenesis and metastasis.
[0198] The compounds of this invention may also be useful in
combination (administered together or sequentially) with known
anti-cancer treatments such as radiation therapy or with cytostatic
or cytotoxic agents, such as for example, but not limited to, DNA
interactive agents; either naturally occurring or synthetic;
hormonal agents, such as tamoxifen; other tyrosine kinase
inhibitors such as Iressa and OSI-774; c-Kit inhibitors.
[0199] The pharmaceutical compositions containing the active
ingredient may be in a form suitable for oral use, for example, as
tablets, troches, lozenges, aqueous or oily suspensions,
dispersible powders or granules, emulsions, hard or soft capsules,
or syrups or elixirs. Compositions intended for oral use may be
prepared according to any method known to the art for the
manufacture of pharmaceutical compositions and such compositions
may contain one or more agents selected from the group consisting
of sweetening agents, flavoring agents, coloring agents and
preserving agents in order to provide pharmaceutically elegant and
palatable preparations. Tablets contain the active ingredient in
admixture with non-toxic pharmaceutically acceptable excipients
which are suitable for the manufacture of tablets. These excipients
may be for example, inert diluents, such as calcium carbonate,
sodium carbonate, lactose, calcium phosphate or sodium phosphate;
granulating and disintegrating agents, for example,
microcrystalline cellulose, sodium crosscarmellose, corn starch, or
alginic acid; binding agents, for example starch, gelatin,
polyvinyl-pyrrolidone or acacia, and lubricating agents, for
example, magnesium stearate, stearic acid or talc. The tablets may
be uncoated or they may be coated by known techniques to mask the
unpleasant taste of the drug or delay disintegration and absorption
in the gastrointestinal tract and thereby provide a sustained
action over a longer period. For example, a water soluble taste
masking material such as hydroxypropyl-methylcellulose or
hydroxypropyl-cellulose, or a time delay material such as ethyl
cellulose, cellulose acetate buryrate may be employed.
[0200] Formulations for oral use may also be presented as hard
gelatin capsules wherein the active ingredient is mixed with an
inert solid diluent, for example, calcium carbonate, calcium
phosphate or kaolin, or as soft gelatin capsules wherein the active
ingredient is mixed with water soluble carrier such as
polyethyleneglycol or an oil medium, for example peanut oil, liquid
paraffin, or olive oil.
[0201] Aqueous suspensions contain the active material in admixture
with excipients suitable for the manufacture of aqueous
suspensions. Such excipients are suspending agents, for example
sodium carboxymethylcellulose, methylcellulose,
hydroxypropylmethyl-cellulose, sodium alginate,
polyvinyl-pyrrolidone, gum tragacanth and gum acacia; dispersing or
wetting agents may be a naturally-occurring phosphatide, for
example lecithin, or condensation products of an alkylene oxide
with fatty acids, for example polyoxyethylene stearate, or
condensation products of ethylene oxide with long chain aliphatic
alcohols, for example heptadecaethyleneoxycetanol, or condensation
products of ethylene oxide with partial esters derived from fatty
acids and a hexitol such as polyoxyethylene sorbitol monooleate, or
condensation products of ethylene oxide with partial esters derived
from fatty acids and hexitol anhydrides, for example polyethylene
sorbitan monooleate. The aqueous suspensions may also contain one
or more preservatives, for example ethyl, or n-propyl
p-hydroxybenzoate, one or more coloring agents, one or more
flavoring agents, and one or more sweetening agents, such as
sucrose, saccharin or aspartame.
[0202] Oily suspensions may be formulated by suspending the active
ingredient in a vegetable oil, for example arachis oil, olive oil,
sesame oil or coconut oil, or in mineral oil such as liquid
paraffin. The oily suspensions may contain a thickening agent, for
example beeswax, hard paraffin or cetyl alcohol. Sweetening agents
such as those set forth above, and flavoring agents may be added to
provide a palatable oral preparation. These compositions may be
preserved by the addition of an anti-oxidant such as butylated
hydroxyanisol or alpha-tocopherol.
[0203] Dispersible powders and granules suitable for preparation of
an aqueous suspension by the addition of water provide the active
ingredient in admixture with a dispersing or wetting agent,
suspending agent and one or more preservatives. Suitable dispersing
or wetting agents and suspending agents are exemplified by those
already mentioned above. Additional excipients, for example
sweetening, flavoring and coloring agents, may also be present.
These compositions may be preserved by the addition of an
anti-oxidant such as ascorbic acid.
[0204] The pharmaceutical compositions of the invention may also be
in the form of an oil-in-water emulsions. The oily phase may be a
vegetable oil, for example olive oil or arachis oil, or a mineral
oil, for example liquid paraffin or mixtures of these. Suitable
emulsifying agents may be naturally-occurring phosphatides, for
example soy bean lecithin, and esters or partial esters derived
from fatty acids and hexitol anhydrides, for example sorbitan
monooleate, and condensation products of the said partial esters
with ethylene oxide, for example polyoxyethylene sorbitan
monooleate. The emulsions may also contain sweetening, flavoring
agents, preservatives and antioxidants.
[0205] Syrups and elixirs may be formulated with sweetening agents,
for example glycerol, propylene glycol, sorbitol or sucrose. Such
formulations may also contain a demulcent, a preservative,
flavoring and coloring agents and antioxidant.
[0206] The pharmaceutical compositions may be in the form of a
sterile injectable aqueous solutions. Among the acceptable vehicles
and solvents that may be employed are water, Ringer's solution and
isotonic sodium chloride solution.
[0207] The sterile injectable preparation may also be a sterile
injectable oil-in-water microemulsion where the active ingredient
is dissolved in the oily phase. For example, the active ingredient
may be first dissolved in a mixture of soybean oil and lecithin.
The oil solution then introduced into a water and glycerol mixture
and processed to form a microemulation.
[0208] The injectable solutions or microemulsions may be introduced
into a patient's blood-stream by local bolus injection.
Alternatively, it may be advantageous to administer the solution or
microemulsion in such a way as to maintain a constant circulating
concentration of the instant compound. In order to maintain such a
constant concentration, a continuous intravenous delivery device
may be utilized. An example of such a device is the Deltec
CADD-PLUS.TM. model 5400 intravenous pump.
[0209] The pharmaceutical compositions may be in the form of a
sterile injectable aqueous or oleagenous suspension for
intramuscular and subcutaneous administration. This suspension may
be formulated according to the known art using those suitable
dispersing or wetting agents and suspending agents which have been
mentioned above. The sterile injectable preparation may also be a
sterile injectable solution or suspension in a non-toxic
parenterally-acceptable diluent or solvent, for example as a
solution in 1,3-butane diol. In addition, sterile, fixed oils are
conventionally employed as a solvent or suspending medium. For this
purpose any bland fixed oil may be employed including synthetic
mono- or diglycerides. In addition, fatty acids such as oleic acid
find use in the preparation of injectables.
[0210] Compounds of Formulas I may also be administered in the form
of a suppositories for rectal administration of the drug. These
compositions can be prepared by mixing the drug with a suitable
non-irritating excipient which is solid at ordinary temperatures
but liquid at the rectal temperature and will therefore melt in the
rectum to release the drug. Such materials include cocoa butter,
glycerinated gelatin, hydrogenated vegetable oils, mixtures of
polyethylene glycols of various molecular weights and fatty acid
esters of polyethylene glycol.
[0211] For topical use, creams, ointments, jellies, solutions or
suspensions, etc., containing the compound of Formula I are
employed. (For purposes of this application, topical application
shall include mouth washes and gargles.)
[0212] The compounds for the present invention can be administered
in intranasal form via topical use of suitable intranasal vehicles
and delivery devices, or via transdermal routes, using those forms
of transdermal skin patches well known to those of ordinary skill
in the art. To be administered in the form of a transdermal
delivery system, the dosage administration will, of course, be
continuous rather than intermittent throughout the dosage regimen.
Compounds of the present invention may also be delivered as a
suppository employing bases such as cocoa butter, glycerinated
gelatin, hydrogenated vegetable oils, mixtures of polyethylene
glycols of various molecular weights and fatty acid esters of
polyethylene glycol.
[0213] When a compound according to this invention is administered
into a human subject, the daily dosage will normally be determined
by the prescribing physician with the dosage generally varying
according to the age, weight, sex and response of the individual
patient, as well as the severity of the patient's symptoms.
[0214] If formulated as a fixed dose, such combination products
employ the compounds of this invention within the dosage range
described above and the other pharmaceutically active agent or
treatment within its approved dosage range. Compounds of Formula I
may also be administered sequentially with known anticancer or
cytotoxic agents when a combination formulation is inappropriate.
The invention is not limited in the sequence of administration;
compounds of Formula I may be administered either prior to or after
administration of the known anticancer or cytotoxic agent(s).
Methods of Preparation
[0215] The compounds of the present invention may be prepared by
methods such as those illustrated in the following schemes.
Solvents, temperatures, pressures, and other reaction conditions
may readily be selected by one of ordinary skill in the art.
Starting materials are commercially available or readily prepared
by one of ordinary skill in the art. These schemes are illustrative
and are not meant to limit the possible techniques one skilled in
the art may use to manufacture compounds disclosed herein.
Different methods may be evident to those skilled in the art.
Additionally, the various steps in the synthesis may be performed
in an alternate sequence or order to give the desired compound(s).
All documents cited herein are incorporated herein by reference in
their entirety.
[0216] In general, the time taken to complete a reaction procedure
will be judged by the person performing the procedure, preferably
with the aid of information obtained by monitoring the reaction by
methods such as HPLC or TLC. A reaction does not have to go to
completion to be useful to this invention. The methods for the
preparation of various heterocycles used to this invention can be
found in standard organic reference books, for example,
"Comprehensive Heterocyclic Chemistry, The Structure, Reactions,
Synthesis and Uses, of Heterocyclic Compounds," Katritzky, A. R.,
Rees, C. W. Eds Pergamon Press New York, First edition 1984, and
"Comprehensive Heterocyclic Chemistry II, A Review of the
Literature 1982-1995: The Structure, Reactions, Synthesis and Uses,
of Heterocyclic Compounds," Katritzky, A. R., Rees, C. W. and
Scriven, E., F. Eds Pergamon Press New York, 1996.
[0217] Unless otherwise specified, the various substituents of the
compounds are defined in the same manner as the formula I compound
of the invention.
[0218] Compounds of Formula I wherein W is CONR.sub.8R.sub.10 where
R.sub.8 is alkyl, cycloalkyl, aryl or heteroaryl and R.sub.10 is H
or small alkyl and R.sub.3 is H; could be made using a general
reaction sequence as depicted in Scheme 1. The corresponding
heterocycle (2H-Q-CO.sub.2R; where R is small alkyl) could be
either purchased commercially or synthesized according to the known
procedures in the literature (Pinto et al, J Med Chem, 566-578
(2001)). The coupling of H-Q-CO.sub.2R with the substituted
benzonitrile (1) could be carried out using alkali metal or
transition metal salt mediated reaction typically carried out in
polar aprotic solvent at 25-200.degree. C. Conversion of cyano to
the amino (3) could be accomplished using chemical or catalytic
reduction (R.sub.2.dbd.H) or addition of appropriate alkylmetal
reagent followed by reduction as described in Synth. Commun.
4067-4075, 28(21) 1998. The free amino group can be combined with
protected alfa-amino acid using standard peptide coupling
conditions, such as BOP-Cl, HBTU or DCC. The ester group R, can be
converted into the corresponding acid derivative (6) using acid or
basic hydrolysis. The acid can be combined with HNR.sub.7R.sub.8
using classical or modified peptide bond formation conditions or
using Lewis acid condition and finally the protective group (PG)
can be removed using appropriate deprotecting conditions.
[0219] Alternatively, the heterocycle containing CO.sub.2R can be
built upon the benzonitrile derivative 8 where L is N or C; using
various known cyclization reactions reported in the literature
(WO6020357 and Bioorg Med Chem Lett, 641 (2001)). This can be taken
forward to the target compound using the conditions as reported in
Scheme 1. ##STR16## ##STR17## Likewise compounds of Formula I
wherein W is COR.sub.6R.sub.8 where R.sub.8 forms a
heterocycloalkyl ring with Q and R.sub.8 is aryl or heteroaryl; can
be synthesized using a general reaction sequence as described
Scheme 3. The required benzonitrile 9 can be synthesized using
procedure as reported in WO2003049681. ##STR18## Compounds of
Formula I wherein W is NR.sub.9R.sub.10 where R.sub.9 is H and
R.sub.10 is COR.sub.11; can be synthesized using a general reaction
sequence as depicted in Scheme 4. Acid 6 can be converted into
corresponding amine using Curtius rearrangement. To obtain amide
derivatives (when R.sub.11=aryl or heteroaryl or cycloalkyl) the
amine can be coupled using acid chloride or fluoride or standard
peptide coupling conditions. To furnish carbamate derivatives (when
R.sub.11.dbd.O-aryl, O-heteroaryl or O-cycloalkyl), the amine can
be combined with corresponding halocarbonate or p-nitrophenyl
carbonate. Corresponding urea (when R.sub.11.dbd.NR.sub.4R.sub.5)
derivatives can be made by coupling the amine with corresponding
isocayanate or N,N-disubstituted carbamyl chloride derivative. The
terminal amine functionality can be unmasked at the end using
appropriate deprotection strategy. ##STR19##
[0220] Compounds of formula I wherein W is a 1,3,4-oxidiazole is
made by a general synthetic route described in Scheme 5. The acid 6
can be combined with acylhydrazide of type
(R.sub.1R.sub.2CONHNH.sub.2) using coupling reagents such as DIC or
PyBrop to yield diacylhydrazide derivative 15. This can be cyclized
to 16 with removal of water using reagents such as DIC or DCC or
Burgess reagent at 60 to 150.degree. C. Finally removal of
protective group such as Boc can be accomplished with mineral or
organic acid. ##STR20##
[0221] Similarly compounds of formula I wherein W is a
1,2,4-oxidiazole is made by a general synthetic route described in
Scheme 6. ##STR21##
[0222] Compounds of formula I wherein W is an aryl fused oxazole or
thiazole or imidazole is made by a general synthetic route
described in Scheme 7. ##STR22##
[0223] Compounds of formula I wherein W is 1,3,4-oxidiazole linked
at 5 position through a nitrogen tether can be made by a general
synthetic route described in Scheme 8. ##STR23## Similarly
compounds of formula I wherein W is 1,3-oxazole can be made by a
general synthetic route described in Scheme 9 ##STR24##
EXAMPLES
1-(3-((2-aminopropanamido)methyl)phenyl)-N-benzyl-3-(trifluoromethyl)-1H-p-
yrazole-5-carboxamide (Example 1)
Example 1
[0224] ##STR25##
Part A: Preparation of
1-(3-(aminomethyl)phenyl)-3-(trifluoromethyl)-1H-pyrazole-5-carboxylic
acid
[0225] To a solution of
1-(3-cyanophenyl)-3-(trifluoromethyl)-1H-pyrazole-5-carboxylic acid
(prepared using procedure reported in J Med Chem, 44, 566 (2001))
(1.40 g, 5.0 mmol) in THF (5 mL) and isopropanol (100 mL) was added
4M HCl in dioxane (5 mL) followed by Pd/C (10% wt, 500 mg) and
PtO.sub.2 (20 mg). The reaction was hydrogenated (55 psi H.sub.2)
for 20 hours and filtered through celite. The filtrate was
concentrated in vacuo. The residue was dissolved in methanol and
passed through SCX column (8.times.5 g), eluted first with methanol
and then 2M NH.sub.3 in methanol. The NH.sub.3 elutants were
combined and concentrated in vacuo. The residue was re-dissolved in
methanol and triethylamine (1 mL) was added. This was concentrated
in vacuo to remove any residual NH.sub.4.sup.+, giving the title
compound as a light yellow solid (1.00 g, 70%).
[0226] MS (ESI) m/z 286.07 (M+H)
Part B: Preparation of 1-(3-((2-(tert
butoxycarbonyl)propanamido)methyl)phenyl)-3-(trifluoromethyl)-1H-pyrazole-
-5-carboxylic acid
[0227] To a suspension of 2 (1.00 g, 3.5 mmol) and triethylamine
(1.0 mL, 7.0 mmol) in CH.sub.2Cl.sub.2 (40 mL) was added
Boc-Ala-OSu (1.10 g, 3.85 mmol). The reaction was stirred at room
temperature for 2 hours, became homogeneous and was concentrated in
vacuo. The residue was dissolved in methanol (4 mL). Water (40 mL)
was added slowly, resulting in a cloudy mixture. To this was added
1.0 M aqueous HCl dropwise while stirring, until pH.about.2. White
precipitate formed and the mixture was filtered. The solid was
washed with water, dissolved in methanol and concentrated in vacuo
to give 3 as a light yellow glass (1.37 g). Another 86 mg of 3 was
recovered from the filtrate when crystals formed after a few days,
giving a total yield of 91% of the title compound. MS (ESI) m/z
457.17 (M+H)
[0228] NMR (CDCl3, .delta., ppm): 10.0 (s, 1H), 8.1 (bs, 1H), 7.9
(s, 1H), 7.7 (d, 1H), 7.5 (d, 1H), 7.37 (dd, 1H), 4.52 (bs, 2H),
4.32 (m, 1H), 1.43 (s, 9H), 1.3 (d, 3H)
Part C: Preparation of 1-(3-((2-aminopropanamido)
methyl)phenyl)-N-benzyl-3-(trifluoromethyl)-1H-pyrazole-5-carboxamide
trifluoroacetic acid salt (Example 1)
[0229] To a solution of HOAt (4.1 mg, 0.03 mmol), EDCI (5.8 mg,
0.03 mmol), and DIEA (0.017 mL, 0.10 mmol) in 1,2-dichloroethane
(0.50 mL) was added 3 (9.1 mg, 0.02 mmol). The reaction was stirred
at room temperature for 30 minutes before benzylamine (0.0055 mL,
0.05 mmol) was added. The reaction was stirred at room temperature
for 20 hours and then heated at 65.degree. C. for 5 hours. The
reaction was concentrated in vacuo. To the residue was added
CH.sub.2Cl.sub.2 (0.6 mL) followed by TFA (0.1 mL). The reaction
was stirred at room temperature for 2 hours, concentrated in vacuo,
azeotroped with methanol and purified by preparative HPLC to give
Example 1 as a white solid (9.5 mg, 85%).
Conditions: Column--YMC ODS (20.times.50 mm)
[0230] Solvents--A--90% water--10% methanol--0.1% TFA [0231] B--10%
water--90% methanol--0.1% TFA [0232] Gradient--25% B to 100% B in
12 min [0233] Retention time: 9 min
[0234] MS (ESI) m/z 446.18 (M+H)
[0235] NMR (CD3OD): 8.2 (s, 1H), 7.96 (s, 1H), 7.71 (d, 1H), 7.33
(m, 5H), 7.2 (m, 1H), 7.1 (d, 1H), 4.6 (s, 1H), 4.4 (s, 1H), 3.9
(m, 1H), 1.3 (d, 3H)
[0236] Preparation of Examples 2 to 85: The following examples were
prepared using a method analogous to that used to prepare Example
1. TABLE-US-00004 TABLE 1 Example # Structure MS m/z 2 ##STR26##
604.6 3 ##STR27## 604.6 4 ##STR28## 529.5 5 ##STR29## 547.5 6
##STR30## 438.47 7 ##STR31## 424.44 8 ##STR32## 432.42 9 ##STR33##
446.45 10 ##STR34## 510.51 11 ##STR35## 460.5 12 ##STR36## 472.5 13
##STR37## 460.5 14 ##STR38## 474.5 15 ##STR39## 460.5 16 ##STR40##
460.5 17 ##STR41## 472.5 18 ##STR42## 472.5 19 ##STR43## 464.4 20
##STR44## 464.4 21 ##STR45## 464.4 22 ##STR46## 482.4 23 ##STR47##
482.4 24 ##STR48## 482.4 25 ##STR49## 482.4 26 ##STR50## 482.4 27
##STR51## 480.9 28 ##STR52## 480.9 29 ##STR53## 460.5 30 ##STR54##
460.5 31 ##STR55## 460.5 32 ##STR56## 514.4 33 ##STR57## 514.4 34
##STR58## 514.4 35 ##STR59## 476.5 36 ##STR60## 476.5 37 ##STR61##
476.5 38 ##STR62## 530.4 39 ##STR63## 530.4 40 ##STR64## 530.4 41
##STR65## 522.5 42 ##STR66## 522.5 43 ##STR67## 538.5 44 ##STR68##
471.5 45 ##STR69## 504.5 46 ##STR70## 524.5 47 ##STR71## 530.6 48
##STR72## 490.5 49 ##STR73## 496.5 50 ##STR74## 515.4 51 ##STR75##
436.4 52 ##STR76## 450.4 53 ##STR77## 452.5 54 ##STR78## 466.5 55
##STR79## 513.5 56 ##STR80## 564 57 ##STR81## 475.4 58 ##STR82##
490.5 59 ##STR83## 488.4 60 ##STR84## 536.5 61 ##STR85## 489.5 62
##STR86## 474.5 63 ##STR87## 551.5 64 ##STR88## 489.5 65 ##STR89##
503.5 66 ##STR90## 534.5 67 ##STR91## 534.5 68 ##STR92## 482.5 69
##STR93## 483.5 70 ##STR94## 483.5 71 ##STR95## 486.5 72 ##STR96##
515.5 73 ##STR97## 439.4 74 ##STR98## 489.5 75 ##STR99## 440.4 76
##STR100## 567.6 77 ##STR101## 495.5 78 ##STR102## 516.5 79
##STR103## 450.4 80 ##STR104## 480.4 81 ##STR105## 512.5 82
##STR106## 436.4 83 ##STR107## 497.5 84 ##STR108## 433.4 85
##STR109## 457.4
N-(3-(5-(5-phenyl-1,3,4-oxadiazol-2-yl)-3(trifluoromethyl)-1H-pyrazol-1-yl-
)benzyl)-2-aminopropanamide (Example 86)
Example 86
[0237] ##STR110##
Part A: Preparation of 1-(3-((2-(tert
butoxycarbonyl)aminopropanamido)methyl)phenyl)-N'-benzoyl-3-(trifluoromet-
hyl)-1H-pyrazole-5-carbohydrazide
[0238] PyBOP (86 mg, 0.16 mmol) was added to a solution of
1-(3-((2-(tert
butoxycarbonyl)propanamido)methyl)phenyl)-3-(trifluoromethyl)-1H-pyrazole-
-5-carboxylic acid (50 mg, 0.11 mmol), benzoic hydrazide (23 mg,
0.16 mmol) and N,N-diisopropylethylamine (21.2 mg, 0.16 mmol) in
dichloromethane (2 mL). The reaction mixture was stirred at room
temperature for 18 hours. The reaction mixture was diluted with
dichloromethane-water (10:2 Ml). The organic layer was washed with
water, dried and concentrated to yield the intermediate which was
carried as it to the next reaction. Diisopropylcarbodiimide (50 mg)
was added to the solution of the intermediate (35 mg, 0.06 mmol) in
DMF (0.1 Ml). The solution was heated at 100.degree. C. for 18
hours. The solution was concentrated in vaccuo and the crude was
purified using preparative HPLC using conditions below to yield
28.3 mg of tert-butyl
1-(3-(5-(5-phenyl-1,3,4-oxadiazol-2-yl)-3-(trifluoromethyl)-1H-pyrazol-1--
yl)benzylamino)-1-oxopropan-2-ylcarbamate.
Conditions: Column--YMC ODS (20.times.50 mm)
[0239] Solvents--A--90% water--10% methanol--0.1% TFA [0240] B--10%
water--90% methanol--0.1% TFA [0241] Gradient--20% B to 100% B in
12 min [0242] Retention time: 10.1 min
[0243] MS (ESI) m/z 557.21 (M+H)
[0244] NMR:8.12(m, 2H), 7.8(s, 1H), 7.6-7.5(m, 4H), 7.2-7.1 (m,
2H), 6.9(s, 1H), 4.5(dd, 2H), 4.2(, 1H), 1.46(s, 9H), 1.3(d,
3H)
Part B: Preparation of
N-(3-(5-(5-phenyl-1,3,4-oxadiazol-2-yl)-3(trifluoromethyl)-1H-pyrazol-1-y-
l)benzyl)-2-aminopropanamide (Example 2)
[0245] tert-butyl
1-(3-(5-(5-phenyl-1,3,4-oxadiazol-2-yl)-3-(trifluoromethyl)-1H-pyrazol-1--
yl)benzylamino)-1-oxopropan-2-ylcarbamate (28.3 mg, 0.05 mmol) was
treated with a mixture of trifluoroacetic acid (0.3 Ml) in
CH.sub.2Cl.sub.2 (3 Ml). The reaction mixture was stirred at room
temperature for 1 hour. The reaction mixture was concentrated in
vacuo. The solution of crude product in methanol was loaded onto a
pre-equilibrated SCX column (3 G in 6 ml cartridge, Waters). The
column was eluted with methanol (2.times.5 Ml) and then with 1 M
ammonia solution in methanol (2.times.5 mL). The fractions
containing methanolic ammonia were concentrated in vacuo to give
pure product (Example 2) (17 mg).
[0246] MS m/z 457.16 (M+H)
[0247] NMR (CD3OD): 7.9 (m, 2H), 7.8 (s, 1H), 7.6 (m, 3H), 7.2 (m,
2H), 7.1 (s, 1H), 4.45 (dd, 2H), 3.86 (m, 1H), 1.3 (d, 3H).
[0248] Preparation of Examples 86 to 164: The following examples
were prepared using a method analogous to that used to prepare
Example 87. TABLE-US-00005 TABLE 2 Compd # Structure MS m/z 87
##STR111## 491.88 88 ##STR112## 471.46 89 ##STR113## 475.42 90
##STR114## 549.53 91 ##STR115## 491.88 92 ##STR116## 475.42 93
##STR117## 458.42 94 ##STR118## 458.42 95 ##STR119## 549.53 96
##STR120## 533.53 97 ##STR121## 487.46 98 ##STR122## 525.43 99
##STR123## 491.88 100 ##STR124## 533.5 101 ##STR125## 471.5 102
##STR126## 482.4 103 ##STR127## 549.5 104 ##STR128## 487.5 105
##STR129## 525.4 106 ##STR130## 471.5 107 ##STR131## 482.4 108
##STR132## 475.4 109 ##STR133## 487.5 110 ##STR134## 525.4 111
##STR135## 471.5 112 ##STR136## 501.4 113 ##STR137## 535.5 114
##STR138## 563.6 115 ##STR139## 515.5 116 ##STR140## 515.5 117
##STR141## 535.5 118 ##STR142## 535.5 119 ##STR143## 507.5 120
##STR144## 507.5 121 ##STR145## 493.4 122 ##STR146## 526.3 123
##STR147## 526.3 124 ##STR148## 526.3 125 ##STR149## 526.3 126
##STR150## 447.4 127 ##STR151## 460.4 128 ##STR152## 463.5 129
##STR153## 477.5 130 ##STR154## 541.5 131 ##STR155## 492.9 132
##STR156## 492.9 133 ##STR157## 472.4 134 ##STR158## 488.4 135
##STR159## 526.4 136 ##STR160## 492.5 137 ##STR161## 517.5 138
##STR162## 508.5 139 ##STR163## 508.5 140 ##STR164## 508.5 141
##STR165## 509.5 142 ##STR166## 513.5 143 ##STR167## 514.5 144
##STR168## 499.5 145 ##STR169## 449.5 146 ##STR170## 463.5 147
##STR171## 463.5 148 ##STR172## 477.5 149 ##STR173## 485.5 150
##STR174## 477.5 151 ##STR175## 477.5 152 ##STR176## 458.4 153
##STR177## 533.5 154 ##STR178## 514.5 155 ##STR179## 501.44 156
##STR180## 464.47 157 ##STR181## 464.47 158 ##STR182## 464.47 159
##STR183## 464.47 160 ##STR184## 472.45 161 ##STR185## 497.45 162
##STR186## 526.42 163 ##STR187## 487.46 164 ##STR188## 423.41 165
##STR189## 578.36
N-(3-(5-(3-phenyl-1,2,4-oxadiazol-5-yl)-3-(trifluoromethyl)-1H-pyrazol-1-y-
l)benzyl)-2-aminopropanamide (Example 166)
Example 166
[0249] ##STR190##
Part A: Preparation of tert-butyl
1-(3-(5-(3-phenyl-1,2,4-oxadiazol-5-yl)-3-(trifluoromethyl)-1H-pyrazol-1--
yl)benzylamino)-1-oxopropan-2-ylcarbamate
[0250] Benzamidoime (16.4 mg, 0.12 mmol) was added to a solution of
1,1'-carbonyldiimidazole (20 mg, 0.12 mmol) and 1-(3-((2-(tert
butoxycarbonyl)propanamido)methyl)phenyl)-3-(trifluoromethyl)-1H-pyrazole-
-5-carboxylic acid (50 mg, 0.1 mmol) in DMF (1 Ml). The reaction
mixture was stirred at room temperature for 4 hours. Additional
1,1'-carbonyldiimidazole (19.5 mg, 0.12 mmol) in DMF (1 Ml) was
added to the reaction mixture. The mixture was heated to
100.degree. C. for 6 hours. The solution was concentrated in vacuo
and the crude product was purified using preparative HPLC to yield
tert-butyl
1-(3-(5-(3-phenyl-1,2,4-oxadiazol-5-yl)-3-(trifluoromethyl)-1H-pyrazol-1--
yl)benzylamino)-1-oxopropan-2-ylcarbamate (2.3 mg).
Conditions: Column--YMC ODS (20.times.50 mm)
[0251] Solvents--A--90% water--10% methanol--0.1% TFA [0252] B--10%
water--90% methanol--0.1% TFA [0253] Gradient--20% B to 100% B in
12 min [0254] Retention time: 10.2 min
[0255] MS (ESI) m/z 557.21 (M+H)
[0256] NMR: 8.1(m, 2H), 7.8(s, 1H), 7.5-7.6(m,4H), 7.1(m, 1H),
7.24(m,1H), 7.0(s,1H), 4.55(m,2H), 4.2(m,1H), 1.48(s,9H),
1.3(d,3H)
Part B: Preparation of
N-(3-(5-(3-phenyl-1,2,4-oxadiazol-5-yl)-3-(trifluoromethyl)-1H-pyrazol-1--
yl)benzyl)-2-aminopropanamide (Example 3)
[0257] tert-butyl
1-(3-(5-(3-phenyl-1,2,4-oxadiazol-5-yl)-3-(trifluoromethyl)-1H-pyrazol-1--
yl)benzylamino)-1-oxopropan-2-ylcarbamate (2.3 mg, 0.004 mmol) was
treated with trifluoroacetic acid (0.1 Ml) in CH.sub.2Cl.sub.2 (1
mL). The reaction mixture was stirred at room temperature for 1
hour. The reaction mixture was concentrated in vacuo. The solution
of crude product in methanol was loaded onto a pre-equilibrated SCX
column (3 G in 6 ml cartridge, Waters). The column was eluted with
methanol (2.times.5 Ml) and then with 1 M ammonia solution in
methanol (2.times.5 mL). The fractions containing methanolic
ammonia were concentrated in vacuo to give title product (Example
3) (1 mg).
[0258] MS (ESI) 457.16 (M+H)
[0259] NMR(CD3OD): 8.1 (d, 2H), 7.8 (s, 1H), 7.6 (m, 3H), 7.0-7.2
(m, 3H), 4.4 (s, 2H), 3.5 (s, 1H), 1.3 (d, 2H).
N-(3-(5-(5-(phenylamino)-1,3,4-oxadiazol-2-yl)-3-(trifluoromethyl)-1H-pyra-
zol-1-yl)benzyl)-2aminopropanamide (Example 167)
Example 167
[0260] ##STR191##
Part A: Preparation of ethyl
1-(3-cyanophenyl)-3-(trifluoromethyl)-1H-pyrazole-5-carboxylate
[0261] A flask was charged with
1-(3-(aminomethyl)phenyl)-3-(trifluoromethyl)-1H-pyrazole-5-carboxylic
acid (5.5 g) in dichloromethane (25 mL), and oxalyl chloride (12
mL) was added to the flask (and two drops of DMF). The solution was
stirred at RT for 1 hour, concentrated in vacuo and re-dissolved in
dichlormethane (10 mL) and ethanol (10 mL). The solution was
stirred at RT for 2 hours. The solution was concentrated in vacuo
and dried by vacuum pump to yield 4.5 G of the title ester.
[0262] MS m/z 310.2 (M+H)
Part B: Preparation of ethyl
1-(3-(aminomethyl)phenyl)-3-(trifluoromethyl)-1H-pyrazole-5-carboxylate
[0263] A flask was charged with ethyl
1-(3-cyanophenyl)-3-(trifluoromethyl)-1H-pyrazole-5-carboxylate
(2.8 g) in ethanol (40 mL) at RT. Palladium on carbon (1.0 g) and
platinum oxide (350 mg), and HCl in water (0.5 mL) was added to the
flask. The reaction solution was stirred at RT with hydrogen gas
from a balloon for 2 hours. The solution was filtered through
celite, washed with methanol (10 mL) and concentrated. The crude
was purified by silica gel column chromatography with 5% methanol
and CH.sub.2Cl.sub.2 giving the pure 1.2 G of product.
[0264] MS m/z 314.19 (M+H)
Part C: Preparation of ethyl
1-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-3-(trifluoro-
methyl)-1H-pyrazole-5-carboxylate
[0265] A flask was charged with ethyl
1-(3-(aminomethyl)phenyl)-3-(trifluoromethyl)-1H-pyrazole-5-carboxylate
(1.20 g) in CH.sub.2Cl.sub.2 (10 mL) at RT. N-Boc-alanine
succinamide (1.1 g) was added to the reaction solution along with
triethylamine (0.53 mL). The reaction solution was stirred at RT
for 2 hours. The solution was concentrated in vaccuo The crude was
purified using silica gel column chromatography (5% ethyl
acetate/heptane) to give 1.7 G of the pure product.
[0266] MS m/z 485 (M+H)
Part D: Preparation of
1-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-3-(trifluoro-
methyl)-1H-pyrazole-5-carboxyhydrazide
[0267] A flask was charged with ethyl
1-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-3-(trifluoro-
methyl)-1H-pyrazole-5-carboxylate (1.78 G), hydrazine monohydrate
(0.445 mL) in ethanol (10 mL). The solution was heated to
60.degree. C. for 6 hours The solution was concentrated in vacuo to
a white powder, aqueous work up with CH.sub.2Cl.sub.2 and H2O,
dried with magnesium sulfate and concentrated giving 1.7 G of the
pure product.
[0268] MS m/z 471 (M+H)
Part E:
tert-butyl-1-(3-(5-(5-(phenylamino)-1,3,4-oxadiazol-2-yl)-3-(trifl-
uoromethyl)-1H-pyrazol-1-yl)benzylamino)-1-oxopropan-2-ylcarbamate
[0269] A vial was charged with
1-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-3-(trifluoro-
methyl)-1H-pyrazole-5-carboxyhydrazide (50 mg),
phenylisothiocyanate (16 mg, 14 .mu.L)) and DMF (3 mL). The
reaction solution was stirred at RT for 2 hours. To the solution
was added 3 ml of DMF and PS-Carbodiimide (PS=polystyrene) (483 mg)
and the solution was heated to 80.degree. C. for 24 hours. To the
solution was added PS-BEMP and P-propylamine and the solution was
shaken at RT for 24 hours. The solution was cooled, filtered and
purified using preparative HPLC using conditions given below [0270]
Conditions: Column--YMC ODS (20.times.50 mm) [0271]
Solvents--A--90% water--10% methanol--0.1% TFA [0272] B--10%
water--90% methanol--0.1% TFA [0273] Gradient--45% B to 100% B in
12 min [0274] Retention time: 10.2 min
[0275] MS m/z 572 (M+H)
[0276] NMR (CD3OD): 7.5-7.1 (m,9H), 6.95 (t, 1H), 4.39 (dd, 2H),
3.94 (m, 1H), 1.45-1.13 (m, 12H)
Part F: Preparation of
N-(3-(5-(5-(phenylamino)-1,3,4-oxadiazol-2-yl)-3-(trifluoromethyl)-1H-pyr-
azol-1-yl)benzyl)-2-aminopropanamide (Example 4)
[0277] A flask was charged with
tert-butyl-1-(3-(5-(5-(phenylamino)-1,3,4-oxadiazol-2-yl)-3-(trifluoromet-
hyl)-1H-pyrazol-1-yl)benzylamino)-1-oxopropan-2-ylcarbamate (31 mg)
in dichloromethane (3 mL) and TFA (0.3 mL). The reaction solution
was stirred at RT for 1 hour. The solvent and excess TFA was
removed in vacuo. The product was converted into the free base by
passing through an SCX column (500 mg) eluting with 2 M ammonia
solution in methanol to give 21 mg of the title product.
[0278] MS m/z 472 (M+H)
[0279] NMR (CD3OD):7.2-7.5(m,9H), 6.96(t,1H), 4.42(dd,2H),
3.88(m,1H), 1.41(d,3H)
[0280] Preparation of Examples 168 to 177: The following examples
were prepared using a method analogous to that used to prepare
Example 166. TABLE-US-00006 TABLE 3 Compd # Structure MS m/z 168
##STR192## 502.47 169 ##STR193## 516.5 170 ##STR194## 532.5 171
##STR195## 532.5 172 ##STR196## 536.92 173 ##STR197## 538.45 174
##STR198## 556.44 175 ##STR199## 578.57 176 ##STR200## 547.47 177
##STR201## 529.52
N-(3-(5-(benzo[d]oxazol-2-yl)-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)--
2-aminopropanamide (Example 178)
Example 178
[0281] ##STR202##
Part A: Preparation of
tert-butyl-1-(3-(5-((2-hydroxyphenyl)carbamoyl)-3-(trifluoromethyl)-1H-py-
razol-1-yl)benzylamino)-1-oxopropan-2-ylcarbamate
[0282] A 10 mL vial was charged with 1-(3-((2-(tert
butoxycarbonyl)propanamido)methyl)phenyl)-3-(trifluoromethyl)-1H-pyrazole-
-5-carboxylic acid (170 mg, 0.37 mmol), PyBrOP (286 mg, 0.55, Hunig
base (0.16 Ml, 0.93 mmol) in 1,4-dioxane (5 mL) followed by
2-aminophenol (60 mg, 0.55 mmol). The solution was heated to
80.degree. C. for 1 hour and then cooled to room temperature and
concentrated in vaccuo. The crude product was purified using silica
gel chromatography (25% ethyl acetate-hexane) to yield 150 mg of
the title compound.
[0283] MS (ESI) m/z 548 (M+H)
Part B: Preparation of
N-(3-(5-(benzo[d]oxazol-2-yl)-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzyl)-
-2-aminopropanamide (Example 5)
[0284] DEAD (0.6 Ml, 0.32 mmol) was added to the stirred suspension
of
tert-butyl-1-(3-(5-((2-hydroxyphenyl)carbamoyl)-3-(trifluoromethyl)-1H-py-
razol-1-yl)benzylamino)-1-oxopropan-2-ylcarbamate (150 mg, 0.27
mmol), PS-triphenylphosphine (190 mg of 3 mmol/g loading) in THF (3
Ml. The reaction mixture was stirred at 85.degree. C. for 10 hours.
The solution was filtered and the filtrate was concentrated in
vaccuo. The crude product was purified using preparative HPLC using
condition shown below [0285] Conditions: Column--YMC ODS
(20.times.50 mm) [0286] Solvents--A--90% water--10% methanol--0.1%
TFA [0287] B--10% water--90% methanol--0.1% TFA [0288]
Gradient--40% B to 100% B in 10 min [0289] Retention time: 10.2
min
[0290] MS m/z 530.1 (M+H)
[0291] The product obtained was dissolved in a mixture of
CH.sub.2Cl.sub.2-TFA (1:0.2 Ml). The reaction mixture was stirred
at room temperature for 1 hour and then concentrated in vaccuo. The
product was converted to free amine by loading the methanolic
solution on a SCX cartridge (500 mg) and eluting with 2M ammonia
solution in methanol. The combined fractions were concentrated in
vaccuo to yield 8 mg of the title product.
[0292] MS m/z 430 (M+H)
[0293] NMR (CD3OD): 7.32-7.7 (m, 9H), 4.5 (s, 2H), 3.7 (m,1H), 1.4
(d,3H).
(2S)--N-(3-(2-methyl-5-(5-phenyl-1,3,4-oxadiazol-2yl)pyrimidin-4-yl)benzyl-
)-2-aminopropanamide (Example 179)
Example 179
[0294] ##STR203##
Part A: Preparation of ethyl
2-(3-bromobenzoyl)-3-(dimethylamino)acrylate
[0295] A mixture containing ethyl 3-(3-bromophenyl)-3-oxopropanoate
(4 G) and N,N-dimethylformamide dimethyl acetal (16 mL) was heated
to 90.degree. C. for 6 h. Reaction mixture was concentrated in
vaccuo to give 6 G of ethyl
2-(3-bromobenzoyl)-3-(dimethylamino)acrylate crude which was used
as such in the next reaction.
Part B: Preparation of ethyl
4-(3-bromophenyl)-2-methylpyrimidine-5-carboxylate
[0296] A solution of sodium ethoxide in ethanol (1.6 Ml, 25%, 6.4
mmol) was added to the suspension of acetaamidine hydrochloride
(0.6 G, 6.4 mmol) in ethanol. The mixture was stirred for 30
minutes. To this mixture was added the solution of ethyl
2-(3-bromobenzoyl)-3-(dimethyl-amino)acrylate (2 G, 1.5 mmol) in
ethanol (8 Ml). The reaction mixture was heated to reflux for 3 h
and then cooled to room temperature and concentrated under vaccuo.
The residue was taken up in a mixture of ethyl acetate-water (60
Ml, 4:1). Organic layer was washed with brine, dried (Na2SO4) and
concentrated to yield the crude which upon silica gel column
chromatographic purification (30% ethyl acetate-hexane) afforded
1.5 G of ethyl
4-(3-bromophenyl)-2-methylpyrimidine-5-carboxylate.
[0297] NMR (CDCl3): 9.05 (s, 1H), 7.74 (t, 1H), 7.62 (dd, 1H), 7.60
(dd, 1H), 7.4 (m, 1H), 4.24 (q, 2H), 2.83 (s, 3H), 1.17 (t, 3H)
Part C: Preparation of
4-(3-bromophenyl)-2-methylpyrimidine-5-carbohydrazide
[0298] Hydrazine hydrate (1 Ml) was added to the solution of ethyl
4-(3-bromophenyl)-2-methylpyrimidine-5-carboxylate (1.5 G) in
ethanol (10 Ml). The reaction mixture was refluxed for 16 h; cooled
at room temperature and concentrated in vaccuo. The residue was
partitioned between dichloromethane-water (50 Ml, 4:1). The organic
layer was washed with water, dried and concentrated to yield 1.4 G
of crude 4-(3-bromophenyl)-2-methylpyrimidine-5-carbohydrazide
which was used as such in the next reaction.
Part D: Preparation of
4-(3-bromophenyl)-2-methyl-5-(5-phenyl-1,3,4-oxadiazol-2-yl)pyrimidine
[0299] PyBroP (0.5 G, 0.9 mmol) was added to the solution of crude
4-(3-bromophenyl)-2-methylpyrimidine-5-carbohydrazide (0.2 G, 0.64
mmol) and N,N-diisopropylethylamine (0.3 Ml) in DMF (1 Ml). The
reaction mixture was stirred at room temperature for 3 h.
Diisopropyl carbodiimide (0.25 G, 0.98 mmol) was added to the
reaction mixture and the mixture was heated to 190.degree. in
microwave reactor for 45 minutes. The reaction mixture was cooled
and partitioned between ethyl acetate-water (50 Ml, 4:1). The
organic layer was washed with brine, dried over Na2SO4 and
concentrated. The crude was purified using silica gel column
chromatography (20% ethyl acetate-hexane) to afford 170 mg of
4-(3-bromophenyl)-2-methyl-5-(5-phenyl-1,3,4-oxadiazol-2-yl)pyrimidine.
[0300] NMR (CDCl3): 9.13 (s, 1H), 8.13 (m, 3H), 7.7 (s,1H), 7.53
(m, 3H), 7.37 (m, 2H), 2.82 (s, 3H)
Part E: Preparation of
3-(2-methyl-5-(5-phenyl-1,3,4-oxadiazol-2-yl)pyrimidin-4-yl)benzonitrile
[0301] Zinc cyanide (30 mg, 0.25 mmol) and ((C6H5)3P)4Pd (0) (10
mg, 0.02 mmol) was added to the solution of
4-(3-bromophenyl)-2-methyl-5-(5-phenyl-1,3,4-oxadiazol-2-yl)pyrimidine
(82 mg, 0.2 mmol) in DMF (2 Ml). The reaction mixture was heated to
195.degree. C. for 45 minutes in a microwave oven. The reaction
mixture was cooled to room temperature and diluted with a mixture
of ethyl acetate-water (25 Ml, 4:1). The organic layer was washed
with brine, dried over Na2SO4 and concentrated in vaccuo to yield
the crude which upon silica gel chromatographic purification
yielded 62 mg of pure
3-(2-methyl-5-(5-phenyl-1,3,4-oxadiazol-2-yl)pyrimidin-4-yl)benzonitrile.
[0302] NMR (CDCl3): 9.13 (s, 1H), 8.32 (m, 1H), 8.13 (m, 2H), 7.7
(s,1H), 7.53 (m, 3H), 7.37 (m, 2H), 2.82 (s, 3H)
Part F: Preparation of
(3-(2-methyl-5-(5-phenyl-1,3,4-oxadiazol-2-yl)pyrimidin-4-yl)phenyl)metha-
namine
[0303] Sodium borohydride (20 mg, 0.54 mmol) was added to the
suspension of
3-(2-methyl-5-(5-phenyl-1,3,4-oxadiazol-2-yl)pyrimidin-4-yl)benzonitri-
le (62 mg, 0.18 mmol) and nickel chloride hexahydrate (52 mg, 0.21
mmol) in ethanol (2 Ml). The reaction mixture was stirred at room
temperature for 1 h and then concentrated in vaccuo. The residue
was partitioned between dichloromethane-water (25 Ml, 4:1). The
organic layer was washed with water, dried and concentrated to
furnish 60 mg of crude
(3-(2-methyl-5-(5-phenyl-1,3,4-oxadiazol-2-yl)pyrimidin-4-yl)phenyl)metha-
namine which was utilized as such in the next reaction.
[0304] MS m/z 344.15 M+H)
Part G: Preparation of
(2S)--N-(3-(2-methyl-5-(5-phenyl-1,3,4-oxadiazol-2yl)pyrimidin-4-yl)benzy-
l)-2-aminopropan-amide
[0305] Boc-S-alanine-OSu (30 mg) was added to the solution of crude
(3-(2-methyl-5-(5-phenyl-1,3,4-oxadiazol-2-yl)pyrimidin-4-yl)phenyl)metha-
namine (60 mg) in dichloromethane (2 Ml). The reaction mixture was
stirred at room temperature for 1 h. To this reaction mixture was
added a trifluoroacetic acid in dichloromethane (2 Ml, 25% v/v).
The resulting mixture was stirred for 45 minutes at room
temperature and concentrated in vacuo. The crude product was
purified using preparative HPLC using conditions below. [0306]
Conditions: Column--YMC ODS (20.times.50 mm) [0307]
Solvents--A--90% water--10% methanol--0.1% TFA [0308] B--10%
water--90% methanol--0.1% TFA [0309] Gradient--25% B to 100% B in
12 min [0310] Retention time: 9.8 min
[0311] MS m/z 415.18 (M+H)
[0312] NMR (CD3OD): 9.2(s, 1H), 7.9 (m, 2H), 7.7 (s,1H), 7.53 (m,
3H), 7.37 (m, 2H), 7.07 (s, 1H), 4.56 (dd, 2H), 3.92 (m, 1H), 2.82
(s, 3H), 1.30 (d, 3H).
(2S)--N-(3-(1-methyl-4-(5-phenyl-1,3,4-oxadiazol-2-yl)-1H-pyrazol-5-yl)ben-
zyl)-2-aminopropanamide (Example 180)
Example 180
[0313] ##STR204##
Part A: Preparation of ethyl
5-(3-cyanophenyl)-1-methyl-1H-pyrazole-4-carboxylate
[0314] A mixture of ethyl-3-cyanobenzoylacetate (1.0 G, 5 mmol) and
N,N-dimethylformamide dimethylacetal (2.74 G, 14 mmol) was heated
to 100.degree. C. for 24 h. The reaction mixture was concentrated
in vacuo and the residue was re-dissolved in methanol (5 Ml),
followed by addition of methyl hydrazine (0.3 Ml, 6 mmol). The
reaction mixture was stirred at room temperature for 2 h and then
concentrated in vaccuo. The residue was partitioned between ethyl
acetate-water (60 Ml, 4:1), washed with water, dried over Na2SO4
and concentrated. The crude was purified using silica gel column
chromatography (1 to 15% ethyl acetate-dichloromethane) to yield
0.36 G of required regioisomer; ethyl
5-(3-cyanophenyl)-1-methyl-1H-pyrazole-4-carboxylate and 0.3 G of
ethyl 3-(3-cyanophenyl)-1-methyl-1H-pyrazole-4-carboxylate.
[0315] MS m/z 256.3(M+H)
Part B: Preparation of ethyl
5-(3-(aminomethyl)phenyl)-1-methyl-1H-pyrazole-4-carboxylate
[0316] 10% Pd on charcoal (100 mg) was added to the solution of
ethyl 5-(3-cyanophenyl)-1-methyl-1H-pyrazole-4-carboxylate (100 mg,
3.9 mmol) and trifluoroacetic acid (50 Ul) in isopropanol. The
reaction mixture was stirred under the blanket of hydrogen gas for
2 h. The reaction mixture was filtered over celite. The filtrate
was concentrated in vaccuo to yield 79 mg of crude (.about.85% by
LCMS) ethyl
5-(3-(aminomethyl)phenyl)-1-methyl-1H-pyrazole-4-carboxylate which
was carried as such to the next reaction.
Part C: Preparation of ethyl
5-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-1-methyl-1H--
pyrazole-4-carboxylate
[0317] Boc-S-alanine-O-succinamide (87 mg, 0.3 mmol) was added to
the solution of ethyl
5-(3-(aminomethyl)phenyl)-1-methyl-1H-pyrazole-4-carboxylate (78.2
mg, 0.3 mmol) and triethylamine (42 Ul) in dichloromethane (2 Ml).
The solution was stirred at room temperature for 45 minutes. The
reaction mixture was diluted with 10 Ml of dichloromethane and 5 mL
of water. The organic layer was washed with water, dried over
Na2SO4 and concentrated to yield the crude which was further
purified by silica gel column chromatography (5% ethyl
acetate-heptane) to afford 89 mg of ethyl
5-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-1-methyl-1H--
pyrazole-4-carboxylate
Part D: Preparation of
5-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-1-methyl-1H--
pyrazole-4-carboxylic acid
[0318] Aqueous sodium hydroxide solution (0.2 Ml, 1 M) was added to
the solution of ethyl
5-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-1-methyl-1H--
pyrazole-4-carboxylate (89 mg) in ethanol (3 Ml) and THF (1 Ml).
The reaction mixture was heated to 60.degree. C. for 6 h. The
reaction was diluted with ethyl acetate-water (10 mL, 1:1). The
organic layer was washed with 1 N NaOH. The combined aqueous layer
was acidified with 1 N HCl and extracted with dichloromethane.
Organic layer was washed with water, dried over Na2SO4 and
concentrated to yield 50 mg of crude
5-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-1-methyl-1H--
pyrazole-4-carboxylic acid which was used as such in the next
reaction.
[0319] MS m/z 403.1(M+H)
Part E: tert-butyl
(S)-1-(3-(1-methyl-4-(5-phenyl-1,3,4-oxadiazol-2-yl)-1H-pyrazol-5-yl)benz-
ylamino)-1-oxopropan-2-ylcarbamate
[0320] Benzoic hydrazide (10.2 mg, 0.07 mmol) was added to the
solution of
5-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-1-methyl--
1H-pyrazole-4-carboxylic acid (50 mg, 0.124 mmol), PyBroP (97 mg,
0.186 mmol) and Hunig's base (54.1 Ul, 0.3 mmol) in DMF (1 Ml). The
reaction mixture was heated at 120.degree. C. for 10 min in
microwave. To this solution was added diisopropylcarbodiimide (100
Ul, 0.621 mmol) and the reaction mixture was further heated to
150.degree. C. for 40 minutes in microwave. The reaction mixture
was diluted with methanol (1 Ml) and the solution was directly
loaded onto a preparative HPLC for purification to yield 25 mg of
tert-butyl
(S)-1-(3-(1-methyl-4-(5-phenyl-1,3,4-oxadiazol-2-yl)-1H-pyrazol-5-yl)benz-
ylamino)-1-oxopropan-2-ylcarbamate.
[0321] The conditions for purification are as follows: [0322]
Conditions: Column--YMC ODS (20.times.50 mm) [0323]
Solvents--A--90% water--10% methanol--0.1% TFA [0324] B--10%
water--90% methanol--0.1% TFA [0325] Gradient--45% B to 100% B in
12 min [0326] Retention time: 12 min
[0327] MS m/z 503.3 (M+H)
[0328] NMR (CDCl3): 8.6 (s, 1H), 8.12 (m, 2H), 7.74 (m, 1H), 7.54
(m, 2H), 7.38 (m, 1H), 7.2 (m, 1H), 7.06 (m, 1H), 6.6 (m, 2H),
4.58-4.67 (m, 2H), 4.3 (m, 1H), 3.7 (s, 3H), 1.5 (s, 9H), 1.3 (d,
3H)
Part F: Preparation of
(2S)--N-(3-(1-methyl-4-(5-phenyl-1,3,4-oxadiazol-2-yl)-1H-pyrazol-5-yl)be-
nzyl)-2-aminopropanamide
[0329] tert-butyl
(S)-1-(3-(1-methyl-4-(5-phenyl-1,3,4-oxadiazol-2-yl)-1H-pyrazol-5-yl)benz-
ylamino)-1-oxopropan-2-ylcarbamate (25 mg) was treated with a
mixture of trifluoroacetic acid in dichloromethane (0.2:1 Ml) at
room temperature for 30 minutes. The reaction mixture was
concentrated in vaccuo and the residue was re-dissolved in methanol
(2 Ml). The methanolic solution was loaded onto a pre-washed and
equilibrated (with methanol) cartridge containing 2 G of SCX
(Waters). The column was eluted with 5 Ml OF METHANOL AND THEN WITH
10 Ml of 2 M ammonia solution in methanol. The later fractions were
concentrated to yield 3 mg of
(2S)--N-(3-(1-methyl-4-(5-phenyl-1,3,4-oxadiazol-2-yl)-1H-pyrazol-5-yl)be-
nzyl)-2-aminopropanamide.
[0330] MS m/z 503.3 (M+H)
[0331] NMR (CD3OD): 8.3(s, 1H), 7.9 (m, 2H), 7.3-7.6 (m, 6H),
4.6(dd, 2H), 3.9 (m, 1H), 3.7 (s, 3H), 1.3 (d, 3H)
(2S)--N-(3-(1-methyl-4-(5-phenyl-1,3,4-oxadiazol-2-yl)-1H-pyrazol-3-yl)ben-
zyl)-2-aminopropanamide (Example 181)
Example 181
[0332] ##STR205##
Part A: Preparation of ethyl
3-(3-cyanophenyl)-1-methyl-1H-pyrazole-4-carboxylate
[0333] Obtained as a minor regioisomer during the preparation of
179 (Part A)
Part B: Preparation of ethyl
3-(3-(aminomethyl)phenyl)-1-methyl-1H-pyrazole-4-carboxylate
[0334] 10% Pd on charcoal (100 mg) was added to the solution of
ethyl 3-(3-cyanophenyl)-1-methyl-1H-pyrazole-4-carboxylate
[0335] (120 mg, 3.9 mmol) and trifluoroacetic acid (50 Ul) in
isopropanol. The reaction mixture was stirred under the blanket of
hydrogen gas for 2 h. The reaction mixture was filtered over
celite. The filtrate was concentrated in vaccuo to yield 79 mg of
crude (.about.85% by LCMS) ethyl
3-(3-(aminomethyl)phenyl)-1-methyl-1H-pyrazole-4-carboxylate which
was carried to the next step without further purification.
[0336] MS m/z 261.2 (M+H)
Part C: Preparation of ethyl
3-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-1-methyl-1H--
pyrazole-4-carboxylate
[0337] Boc-S-alanine-O-succinamide (87 mg, 0.3 mmol) was added to
the solution of ethyl
3-(3-(aminomethyl)phenyl)-1-methyl-1H-pyrazole-4-carboxylate (79
mg, 0.3 mmol) and triethylamine (42 Ul) in dichloromethane (2 Ml).
The solution was stirred at room temperature for 45 minutes. The
reaction mixture was diluted with 10 Ml of dichloromethane and 5 mL
of water. The organic layer was washed with water, dried over
Na2SO4 and concentrated to yield the crude which was further
purified by silica gel column chromatography (5% ethyl
acetate-heptane) to afford 89 mg of ethyl
5-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-1-methyl-1H--
pyrazole-4-carboxylate.
[0338] MS m/z 431.3(M+H)
Part D:
3-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-1-met-
hyl-1H-pyrazole-4-carboxylic acid hydrazide
[0339] Hydrazine monohydrate (23 uL) was added to the solution of
ethyl
5-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-1-methyl-1H--
pyrazole-4-carboxylate (77 mg, 0.18 mmol) in ethanol. The solution
was heated for 5 days at 60.degree. C. and then cooled to room
temperature and concentrated in vaccuo. The residue was partitioned
between water and dichloromethane (25 Ml, 1:4). The organic layer
was washed with water, dried and concentrated to yield 46 mg of
3-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-1-methyl-1H--
pyrazole-4-carboxylic acid hydrazide.
[0340] MS m/z 417(M+H)
Part E: Preparation of tert-butyl
(S)-1-(3-(1-methyl-4-(5-phenyl-1,3,4-oxadiazol-2-yl)-1H-pyrazol-3-yl)benz-
ylamino)-1-oxopropan-2-ylcarbamate
[0341] PyBrOP (84.5 mg, 0.17 mmol) was added to the solution of
3-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-1-methyl-1H--
pyrazole-4-carboxylic acid hydrazide (45 mg, 0.11 mmol), benzoic
acid (20 mg, 0.17 mmol) and Hunig base (47 Ul, 0.27 mmol) in DMF (1
Ml). The reaction mixture was heated at 120.degree. C. in microwave
for 10 minutes. The reaction mixture was cooled to room temperature
and di-isopropylcarbodiimide (85 mg, 0.54) was added. The reaction
mixture was heated at 150.degree. C. for 30 minutes in microwave
oven. The reaction was cooled to room temperature and the crude was
purified using preparative HPLC using the conditions below to
afford 5.2 mg of tert-butyl
(S)-1-(3-(1-methyl-4-(5-phenyl-1,3,4-oxadiazol-2-yl)-1H-pyrazol-3-yl)benz-
ylamino)-1-oxopropan-2-ylcarbamate. [0342] Conditions: Column--YMC
ODS (20.times.50 mm) [0343] Solvents--A--90% water--10%
methanol--0.1% TFA [0344] B--10% water--90% methanol--0.1% TFA
[0345] Gradient--45% B to 100% B in 12 min [0346] Retention time:
12 min
[0347] MS m/z 503.3 (M+H)
[0348] NMR (CDCl3): 8.6 (s, 1H), 8.12 (m, 2H), 7.74 (m, 1H), 7.54
(m, 2H), 7.38 (m, 1H), 7.2 (m, 1H), 7.06 (m, 1H), 6.6 (m, 2H),
4.58-4.67 (m, 2H), 4.3 (m, 1H), 3.7 (s, 3H), 1.5 (s, 9H), 1.3 (d,
3H)
Part F: Preparation of
(2S)--N-(3-(1-methyl-4-(5-phenyl-1,3,4-oxadiazol-2-yl)-1H-pyrazol-3-yl)be-
nzyl)-2-aminopropanamide
[0349] tert-butyl
(S)-1-(3-(1-methyl-4-(5-phenyl-1,3,4-oxadiazol-2-yl)-1H-pyrazol-3-yl)benz-
ylamino)-1-oxopropan-2-ylcarbamate was treated with a mixture of
trifluoroacetic acid in dichloromethane (0.2:1 Ml) at room
temperature for 30 minutes. The reaction mixture was concentrated
in vaccuo and the residue was re-dissolved in methanol (2 Ml). The
methanolic solution was loaded onto a pre-washed and equilibrated
(with methanol) cartridge containing 2 G of SCX (Waters). The
column was eluted with 5 Ml OF METHANOL AND THEN WITH 10 Ml of 2 M
ammonia solution in methanol. The later fractions were concentrated
to yield 3 mg of
(2S)--N-(3-(1-methyl-4-(5-phenyl-1,3,4-oxadiazol-2-yl)-1H-pyrazol-5-yl)be-
nzyl)-2-aminopropanamide.
[0350] MS m/z 503.3 (M+H)
[0351] NMR (CD3OD): 8.3(s, 1H), 7.9 (m, 2H), 7.3-7.6 (m, 6H),
4.6(dd, 2H), 3.9 (m, 1H), 3.7 (s, 3H), 1.3 (d, 3H)
(2S)--N-(3-(5-(5-phenyl-1,3,4-oxadiazol-2-yl)-1H-imidazol-1-yl)benzyl)-2-a-
minopropanamide (Example 182)
Example 182
[0352] ##STR206##
Part A: Preparation of methyl
1-(3-cyanophenyl)-1H-imidazole-5-carboxylate
[0353] Using the procedure as described in example #191 in U.S.
Pat. No. 6,548,512
Part B: Preparation of methyl
1-(3-(aminomethyl)phenyl)-1H-imidazole-5-carboxylate
[0354] 10% Pd on charcoal (200 mg) and platinum oxide (100 mg) were
added to the solution of methyl
1-(3-cyanophenyl)-1H-imidazole-5-carboxylate (300 mg, 1.32 mmol)
and trifluoroacetic acid (0.1 mL) in ethanol. The reaction mixture
was stirred under the blanket of hydrogen gas for 24 h. The
reaction mixture was filtered over celite. The filtrate was
concentrated in vaccuo to give the crude. The crude was purified
using preparative HPLC using conditions below to yield 100 mg of
methyl 1-(3-(aminomethyl)phenyl)-1H-imidazole-5-carboxylate [0355]
Conditions: Column--YMC ODS (20.times.50 mm) [0356]
Solvents--A--90% water--10% methanol--0.1% TFA [0357] B--10%
water--90% methanol--0.1% TFA [0358] Gradient--25% B to 100% B in
12 min [0359] Retention time: 9.6 min
[0360] MS m/z 232.2 (M+H)
Part C: Preparation of methyl
1-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-1H-imidazole-
-5-carboxylate
[0361] Boc-S-alanine-O-succinamide (125 mg, 0.43 mmol) was added to
the solution of methyl
1-(3-(aminomethyl)phenyl)-1H-imidazole-5-carboxylate (101 mg, 0.43
mmol) and triethylamine (60 Ul) in dichloromethane (2 Ml). The
solution was stirred at room temperature for 45 minutes. The
reaction mixture was diluted with 10 Ml of dichloromethane and 5 mL
of water. The organic layer was washed with water, dried over
Na2SO4 and concentrated to yield the crude which was further
purified by silica gel column chromatography (5% ethyl
acetate-heptane) to afford 89 mg of ethyl
5-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-1-methyl-1H--
pyrazole-4-carboxylate.
[0362] NMR (CDCl3): 7.6 (s, 1H), 7.3-7.5 (m, 3H), 7.2 (m, 1H), 7.06
(s, 1H), 6.1 (m, 2H), 4.67 (m, 2H), 4.1 (m, 1H), 3.7 (s, 3H),
1.3-1.5 (m, 12H)
[0363] MS m/z 431.3(M+H)
Part D: Preparation of
1-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-1H-imidazole-
-5-carboxylic acid hydrazide
[0364] Hydrazine monohydrate (14 uL) was added to the solution of
methyl 1-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)
methyl)phenyl)-1H-imidazole-5-carboxylate (46 mg, 0.113 mmol) in
ethanol (5 Ml). The solution was heated for 24 h at 60.degree. C.
and then cooled to room temperature and concentrated in vaccuo. The
residue was partitioned between water and dichloromethane (25 Ml,
1:4). The organic layer was washed with water, dried and
concentrated to yield 32 mg of
1-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)
methyl)phenyl)-1H-imidazole-5-carboxylic acid hydrazide.
[0365] MS m/z 403(M+H)
Part E: Preparation of tert-butyl
(S)-1-(3-(5-(5-phenyl-1,3,4-oxadiazol-2-yl)-1H-imidazol-1-yl)benzylamino)-
-1-oxopropan-2-ylcarbamate
[0366] PyBrOP (62.5 mg, 0.12 mmol) was added to the solution of
1-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-1H-imidazole-
-5-carboxylic acid hydrazide (32 mg, 0.08 mmol), benzoic acid (10
mg, 0.08 mmol) and Hunig base (35 Ul, 0.2 mmol) in DMF (1 Ml). The
reaction mixture was heated at 120.degree. C. in microwave for 15
minutes. The reaction mixture was cooled to room temperature and
di-isopropylcarbodiimide (62 mg, 0.40) was added. The reaction
mixture was heated at 150.degree. C. for 30 minutes in microwave
oven. The reaction was cooled to room temperature and the crude was
purified using preparative HPLC using the conditions below to
afford 40 mg of tert-butyl
(S)-1-(3-(5-(5-phenyl-1,3,4-oxadiazol-2-yl)-1H-imidazol-1-yl)benzylamino)-
-1-oxopropan-2-ylcarbamate. [0367] Conditions: Column--YMC ODS
(20.times.50 mm) [0368] Solvents--A--90% water--10% methanol--0.1%
TFA [0369] B--10% water--90% methanol--0.1% TFA [0370]
Gradient--45% B to 100% B in 12 min [0371] Retention time: 10
min
[0372] MS m/z 489 (M+H)
[0373] NMR (CDCl3): 8.6 (s, 1H), 8.12 (m, 2H), 7.74 (m, 1H), 7.54
(m, 2H), 7.38 (m, 1H), 7.2 (m, 1H), 7.06 (m, 1H), 6.6 (m, 2H),
4.58-4.67 (m, 2H), 4.3 (m, 1H), 3.7 (s, 3H), 1.5 (s, 9H), 1.3 (d,
3H)
Part F: Preparation of
(2S)--N-(3-(5-(5-phenyl-1,3,4-oxadiazol-2-yl)-1H-imidazol-1-yl)benzyl)-2--
aminopropanamide trifluoroacetate salt
[0374] 39 mg (0.08 mmol) of tert-butyl
(S)-1-(3-(5-(5-phenyl-1,3,4-oxadiazol-2-yl)-1H-imidazol-1-yl)benzylamino)-
-1-oxopropan-2-ylcarbamate in dichloromethane (2 Ml) was treated
trifluoroacetic acid (0.4 Ml). The reaction mixture was stirred at
room temperature for 45 minutes and then concentrated in vaccuo to
afford 20 mg of
(2S)--N-(3-(5-(5-phenyl-1,3,4-oxadiazol-2-yl)-1H-imidazol-1-yl)benz-
yl)-2-aminopropanamide trifluoroacetate salt.
[0375] MS m/z 389 (M+H)
[0376] NMR (CD3OD): 8.4 (s, 1H), 8.12 (m, 1H), 7.92 (d, 2H), 7.54
(m, 5H), 4.50(dd, 2H), 3.95 (m, 1H), 1.3 (d, 3H)
(2S)--N-(3-(4-(5-phenyl-1,3,4-oxadiazol-2-yl)oxazol-5-yl)benzyl)-2-aminopr-
opanamide (Example 183)
Example 183
[0377] ##STR207##
Part A: Preparation of methyl
5-(3-cyanophenyl)oxazole-4-carboxylate
[0378] Methyl isocyanoacetate (0.5 Ml, 6 mmol) was added to the
solution of 3-cyanobenzoyl chloride (1 G, 6 mmol) and triethylamine
(2.5 Ml) in dichloromethane (20 ml). The reaction mixture was
stirred for 24 h. The reaction mixture was diluted with 20 mL of
dichloromethane and 10 mL of water. The organic layer was washed
with water, dried over Na2SO4 and concentrated. The crude product
was recrystallized from hot methanol to give 0.98 G of methyl
5-(3-cyanophenyl)oxazole-4-carboxylate
[0379] NMR (CDCl3):8.7 (s, 1H), 8.2 (d, 1H), 7.93 (s, 1H), 7.82 (d,
1H), 7.6 (m, 1H), 3.78 (s, 3H)
Part B: Preparation of methyl
5-(3-(aminomethyl)phenyl)oxazole-4-carboxylate
[0380] 10% Pd on charcoal (100 mg) and platinum oxide (170 mg) were
added to the solution of methyl
1-(3-cyanophenyl)-1H-imidazole-5-carboxylate (300 mg, 1.32 mmol)
and trifluoroacetic acid (50 uL) in isopropanol. The reaction
mixture was stirred under the blanket of hydrogen gas for 4 h. The
reaction mixture was filtered over celite. The filtrate was
concentrated in vaccuo to give the crude. The crude was dissolved
in methanol and loaded over a SCX column (2 G) pre-equilibrated
with methanol. The column was eluted with 10 Ml of methanol and
then 10 mL of 2N methanolic ammonia solution. The fractions from
the later elution were combined and concentrated to yield 123 mg of
methyl 5-(3-(aminomethyl)phenyl)oxazole-4-carboxylate.
[0381] MS m/z 234 (M+H)
Part C: Preparation of methyl
5-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)oxazole-4-car-
boxylate
[0382] Boc-S-alanine-O-succinamide (153 mg, 0.53 mmol) was added to
the solution of methyl
5-(3-(aminomethyl)phenyl)oxazole-4-carboxylate (123 mg, 0.53 mmol)
and triethylamine (75 uL) in dichloromethane (2 mL). The solution
was stirred at room temperature for 45 minutes. The reaction
mixture was diluted with 10 Ml of dichloromethane and 5 mL of
water. The organic layer was washed with water, dried over Na2SO4
and concentrated to yield the crude which was further purified by
silica gel column chromatography (5% ethyl acetate-heptane) to
afford 106 mg of methyl
5-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)oxazole-4-car-
boxylate
[0383] NMR (CDCl3): 8.7 (s, 1H), 7.8 (d, 1H), 7.35 (m, 2H), 7.15
(d, 1H), 6.1 (m, 1H), 4.7 (s, 2H), 4.1 (m, 1H), 3.8 (s, 3H), 1.4
(m, 12H)
[0384] MS m/z 404(M+H)
Part D: Preparation of
5-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)oxazole-4-car-
boxylic acid hydrazide
[0385] Hydrazine monohydrate (30 uL) was added to the solution of
methyl 5-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)
methyl)phenyl)oxazole-4-carboxylate (106 mg, 0.26 mmol) in ethanol
(5 mL). The solution was heated for 1 h at 60.degree. C. and then
cooled to room temperature and concentrated in vaccuo. The residue
was partitioned between water and dichloromethane (25 mL, 1:4). The
organic layer was washed with water, dried over Na2SO4 and
concentrated to yield 91 mg of
5-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)
methyl)phenyl)oxazole-4-carboxylic acid hydrazide.
[0386] MS m/z 404(M+H)
Part E: Preparation of tert-butyl
(S)-1-(3-(4-(5-phenyl-1,3,4-oxadiazol-2-yl)oxazol-5-yl)benzylamino)-1-oxo-
propan-2-ylcarbamate
[0387] PyBrOP (174 mg, 0.336 mmol) was added to the solution of
5-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)
methyl)phenyl)oxazole-4-carboxylic acid hydrazide (90 mg, 0.224
mmol), benzoic acid (41 mg, 0.336 mmol) and Hunig base (98 uL, 0.56
mmol) in DMF (1 Ml). The reaction mixture was heated at 120.degree.
C. in microwave for 15 minutes. The reaction mixture was cooled to
room temperature and di-isopropylcarbodiimide (141 mg, 1.12) was
added. The reaction mixture was heated at 150.degree. C. for 30
minutes in microwave oven. The reaction was cooled to room
temperature and the crude was purified using preparative HPLC using
the conditions below to afford 40 mg of tert-butyl
(S)-1-(3-(4-(5-phenyl-1,3,4-oxadiazol-2-yl)oxazol-5-yl)benzylamino)-1-oxo-
propan-2-ylcarbamate. [0388] Conditions: Column--YMC ODS
(20.times.50 mm) [0389] Solvents--A--90% water--10% methanol--0.1%
TFA [0390] B--10% water--90% methanol--0.1% TFA [0391]
Gradient--45% B to 100% B in 12 min [0392] Retention time: 10
min
[0393] NMR (CDCl3):8.21 (s, 1H), 8.1 (m, 2H), 7.7 (d, 1H), 7.5 (m,
3H), 7.33 (m, 2H), 7.0 (d, 1H), 6.8 (bs, 1H), 4.63 (dd, 2H), 4.32
(m, 1H), 1.5 (s, 9H), 1.32 (d, 3H)
[0394] MS m/z 490 (M+H)
Part F: Preparation of
(2S)--N-(3-(4-(5-phenyl-1,3,4-oxadiazol-2-yl)oxazol-5-yl)benzyl)-2-aminop-
ropanamide
[0395] Trifluoroacetic acid (1 Ml) was added to the solution of
tert-butyl
(S)-1-(3-(4-(5-phenyl-1,3,4-oxadiazol-2-yl)oxazol-5-yl)benzylamino)-1-oxo-
propan-2-ylcarbamate (32 mg) in dichloromethane (3 Ml). The
reaction mixture was stirred for 45 minutes at room temperature and
then concentrated in vaccuo. The product was dissolved in 2 mL of
methanol and loaded onto pre-equilibrated (with methanol) column of
SCX. The column was eluted with methanol (2.times.2 mL) and then
with 2 N solution of ammonia in methanol. The fractions were
concentrated to yield 16 mg of
(2S)--N-(3-(4-(5-phenyl-1,3,4-oxadiazol-2-yl)oxazol-5-yl)benzyl)-2-aminop-
ropanamide.
[0396] MS m/z 390.3 (M+H)
[0397] NMR (CD3OD): 8.6 (s, 1H), 8.4 (s, 1H), 8.2 (m, 3H), 7.6 (m,
7H), 4.5 (dd, 2H), 3.6 (m, 1H), 1.32 (d, 3H)
(2S)--N-(3-(5-methyl-4-(5-phenyl-1,3,4-oxadiazol-2-yl)isoxazol-3-yl)benzyl-
)-2-aminopropanamide (Example 184)
Example 184
[0398] ##STR208##
Part A: Preparation of
3-(3-(aminomethyl)phenyl)-5-methylisoxazole-4-carboxylic acid
[0399] Using the procedure as reported in WO 9828282
Part B: Preparation of
3-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-5-methylisox-
azole-4-carboxylic acid
[0400] Boc-S-alanine-O-succinamide (94 mg, 0.33 mmol) was added to
the solution of
3-(3-(aminomethyl)phenyl)-5-methylisoxazole-4-carboxylic acid (70
mg, 0.3 mmol) and triethylamine (75 uL) in dichloromethane (2 mL).
The solution was stirred at room temperature for 45 minutes. The
reaction mixture was diluted with 10 Ml of dichloromethane and 5 mL
of water. The organic layer was washed with water, dried over
Na2SO4 and concentrated to yield the crude which was further
purified by silica gel column chromatography (5% ethyl
acetate-heptane) to afford 120 mg of
3-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-5-methylisox-
azole-4-carboxylic acid
[0401] NMR (CDCl3): 11 (s, 1H), 7.2-7.3 (m, 3H), 7.15 (d, 1H), 4.5
(s, 2H), 3.4 (m, 1H), 1.4 (m, 12H),
[0402] MS m/z 404(M+H)
Part B: Preparation of tert-butyl
(S)-1-(3-(5-methyl-4-(5-phenyl-1,3,4-oxadiazol-2-yl)isoxazol-3-yl)benzyla-
mino)-1-oxopropan-2-ylcarbamate
[0403] PyBroP (44 mg, 0.08 mmol) was added to the solution of
3-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-5-methylisox-
azole-4-carboxylic acid (28 mg, 0.07 mmol) and
N,N-diisopropylethylamine (18 mg, 0.14 mmol) in DMF (1 Ml). The
reaction mixture was stirred for 16 h at room temperature and then
di-isopropylcarbodiimide (36 mg, 0.28 mmol) was added to the
reaction mixture. The reaction mixture was heated at 180.degree. C.
in a microwave oven for 30 minutes. The reaction mixture was
diluted with 1 mL of methanol and the crude was purified using
preparative HPLC using the conditions as below to yield 7 mg of
tert-butyl
(S)-1-(3-(5-methyl-4-(5-phenyl-1,3,4-oxadiazol-2-yl)isoxazol-3-yl)benzyla-
mino)-1-oxopropan-2-ylcarbamate [0404] Conditions: Column--YMC ODS
(20.times.50 mm) [0405] Solvents--A--90% water--10% methanol--0.1%
TFA [0406] B--10% water--90% methanol--0.1% TFA [0407]
Gradient--45% B to 100% B in 12 min [0408] Retention time: 9.5
min
[0409] MS m/z 504 (M+H)
Part C: Preparation of
(2S)--N-(3-(5-methyl-4-(5-phenyl-1,3,4-oxadiazol-2-yl)isoxazol-3-yl)benzy-
l)-2-aminopropanamide
[0410] 7 mg of tert-butyl
(S)-1-(3-(5-methyl-4-(5-phenyl-1,3,4-oxadiazol-2-yl)isoxazol-3-yl)benzyla-
mino)-1-oxopropan-2-ylcarbamate was treated with 0.1 mL of
trifluoroacetic acid in 0.5 mL dichloromethane. The reaction
mixture was stirred at room temperature for 30 minutes and then
evaporated in vaccuo to yield 6 mg of trifluoroacetic acid salt of
(2S)--N-(3-(5-methyl-4-(5-phenyl-1,3,4-oxadiazol-2-yl)isoxazol-3-yl)benzy-
l)-2-aminopropanamide.
[0411] NMR (CD3OD): 7.89 (m, 2H), 7.74 (s, 1H), 7.66-7.51 (m, 6H),
4.51 (m, 2H), 3.98 (q, 1H), 2.87 (s, 3H), 1.51 (d, 3H)
[0412] MS m/z 404 (M+H)
N-(3-(3-methyl-5-(5-phenyl-1,3,4-oxadiazol-2-yl)-1H-pyrazol-1-yl)benzyl)-2-
-aminopropanamide (Example 185)
Example 185
[0413] ##STR209##
Part A: Preparation of
1-(3-(aminomethyl)phenyl)-3-methyl-1H-pyrazole-5-carboxylic acid
hydrate
[0414] Using the procedure as reported (Pinto et al, J Med Chem,
566-578 (2001)).
Part B: Preparation of
1-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-3-methyl-1H--
pyrazole-5-carboxylic acid
[0415] Boc-S-alanine-O-succinamide (94 mg, 0.33 mmol) was added to
the solution of
3-(3-(aminomethyl)phenyl)-5-methylisoxazole-4-carboxylic acid (70
mg, 0.3 mmol) and triethylamine (152 uL) in dichloromethane (2 mL).
The solution was stirred at room temperature for 45 minutes. The
reaction mixture was diluted with 10 mL of dichloromethane and 5 mL
of water. The organic layer was washed with water, dried over
Na2SO4 and concentrated to yield the crude which was further
purified by silica gel column chromatography (5% ethyl
acetate-heptane) to afford 120 mg of
3-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-5-methylisox-
azole-4-carboxylic acid
[0416] NMR (CDCl3): 11 (s, 1H), 7.2-7.3 (m, 3H), 7.15 (d, 1H), 4.5
(s, 2H), 3.4 (m, 1H), 1.4 (m, 12H),
[0417] MS m/z 404(M+H)
Part C: Preparation of
N-(3-(3-methyl-5-(5-phenyl-1,3,4-oxadiazol-2-yl)-1H-pyrazol-1-yl)benzyl)--
2-aminopropanamide (TFA salt)
[0418] PyBroP (44 mg, 0.084 mmol) was added to the solution of
3-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-5-methylisox-
azole-4-carboxylic acid (28 mg, 0.07 mmol), benzoic hydrazide (11.4
mg, 0.084 mmol) and N,N-diisopropylethylamine (25 Ul, 0.14 mmol) in
DMF (0.5 Ml). The reaction mixture was stirred at room temperature
for 2 h. Diisopropylcarbodiimide (36 mg, 0.28 mmol) was added to
the reaction mixture and heated in a microwave oven at 185.degree.
C. for 30 minutes. The reaction mixture was directly loaded onto a
preparative HPLC column equilibrated with 25% solvent B (as defined
below) and purified using the conditions below: [0419] Conditions:
Column--YMC ODS (20.times.50 mm) [0420] Solvents--A--90% water--10%
methanol--0.1% TFA [0421] B--10% water--90% methanol--0.1% TFA
[0422] Gradient--25% B to 100% B in 12 min [0423] Retention time:
10 min
[0424] MS m/z 503.3 (M+H)
[0425] The product was taken up in a mixture of dichloromethane and
trifluoroacetic acid (0.8:0.2 Ml) and stirred at room temperature
for 30 minutes. The solvent was evaporated in vaccuo to afford 7.6
mg of trifluoroacetic acid salt of
N-(3-(3-methyl-5-(5-phenyl-1,3,4-oxadiazol-2-yl)-1H-pyrazol-1-yl)benzyl)--
2-aminopropanamide
[0426] NMR(CD.sub.3OD) 7.92 (m, 2H), 7.65-7.45 (m, 7H), 7.12 (s,
1H), 4.53 (m, 2H), 3.97 (q, J=7.1 Hz, 1H), 2.43 (s, 3H), 1.51 (d,
J=7.1 Hz, 3H).
[0427] MS m/z 517.3 (M+H)
(2S)-2-amino-N-((2-methyl-5-(7-oxo-6-(4-(2-oxopiperidin-1-yl)phenyl)-3-(tr-
ifluoromethyl)-4,5,6,7-tetrahydropyrazolo[3,4-c]pyridin-1-yl)phenyl)methyl-
)propanamide (Example 186)
Example 186
[0428] ##STR210##
Part A: Preparation of
1-(3-(aminomethyl)-4-methylphenyl)-6-(4-(2-oxopiperidin-1-yl)phenyl)-3-(t-
rifluoromethyl)-5,6-dihydro-1H-pyrazolo[3,4-c]pyridin-7(4H)-one
[0429] Using the same procedure as reported in WO2003049681
Part B: Preparation of
1-(3-(aminomethyl)-4-methylphenyl)-6-(4-(2-oxopiperidin-1-yl)phenyl)-3-(t-
rifluoromethyl)-5,6-dihydro-1H-pyrazolo[3,4-c]pyridin-7(4H)-one
[0430] Boc-S-alanine-O-succinamide (31 mg, 0.11 mmol) was added to
the solution of
1-(3-(aminomethyl)-4-methylphenyl)-6-(4-(2-oxopiperidin-1-yl)phenyl)-3-(t-
rifluoromethyl)-5,6-dihydro-1H-pyrazolo[3,4-c]pyridin-7(4H)-one (30
mg, 0.1 mmol) and triethylamine (75 uL) in dichloromethane (2 mL).
The solution was stirred at room temperature for 45 minutes. The
reaction mixture was diluted with 10 Ml of dichloromethane and 5 mL
of water. The organic layer was washed with water, dried over
Na2SO4 and concentrated to yield the crude.
[0431] The crude was treated with a mixture of trifluoroacetic acid
and dichloromethane (0.2:1 Ml) for 45 minutes. The reaction mixture
was concentrated to give the crude which was purified using
preparative HPLC using the conditions outlined below to afford 8.5
mg of trifluoroacetic acid salt of
1-(3-(aminomethyl)-4-methylphenyl)-6-(4-(2-oxopiperidin-1-yl)phenyl)-3-(t-
rifluoromethyl)-5,6-dihydro-1H-pyrazolo[3,4-c]pyridin-7(4H)-one
[0432] Conditions: Column--YMC ODS (20.times.50 mm) [0433]
Solvents--A--90% water--10% methanol--0.1% TFA [0434] B--10%
water--90% methanol--0.1% TFA [0435] Gradient--25% B to 100% B in
12 min [0436] Retention time: 8.8 min
[0437] MS m/z 569.5(M+H)
Benzyl
1-(3-(((S)-2-aminopropanamido)methyl)phenyl)-3-(trifluoromethyl)-1H-
-pyrazol-5-ylcarbamate (Example 187)
Example 187
[0438] ##STR211##
Part A: Preparation of benzyl
1-(3-(((S)-2-(tert-butoxycarbonyl)aminopropanamido)methyl)phenyl)-3-(trif-
luoromethyl)-1H-pyrazol-5-ylcarbamate
[0439] Diphenylphosphoryl azide (19 mg, 0.07 mmol) was added to the
solution of
1-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-3-(trifluoro-
methyl)-1H-pyrazole-5-carboxylic acid (23 mg, 0.05 mmol) and
triethylamine (10 mg, 0.1 mmol) in 1,4-dioxane (0.6 Ml). The
reaction mixture was stirred at room temperature for 16 h. Benzyl
alcohol (22 mg, 0.2 mmol) was added to the reaction and the mixture
was heated at 80.degree. C. for 1 h. The reaction mixture was
concentrated and the crude was purified using silica gel column
chromatography (1% MeOH--CH.sub.2Cl.sub.2) to afford 22 mg of
benzyl
1-(3-(((S)-2-(tert-butoxycarbonyl)aminopropanamido)methyl)phenyl)-3-(trif-
luoromethyl)-1H-pyrazol-5-ylcarbamate
[0440] MS m/z 562.6 (M+H)
Part B: Preparation of benzyl
1-(3-(((S)-2-aminopropanamido)methyl)phenyl)-3-(trifluoromethyl)-1H-pyraz-
ol-5-ylcarbamate
[0441] Benzyl 1-(3-(((S)-2-(tert butoxycarbonyl)aminopropanamido)
methyl)phenyl)-3-(trifluoromethyl)-1H-pyrazol-5-ylcarbamate (9 mg,
0.016 mmol) was treated with a mixture of trifluoroacetic acid in
dichloromethane (0.1:0.3 Ml). The reaction mixture was stirred at
room temperature for 45 minutes and then concentrated in vaccuo to
yield 9 mg of trifluoroacetic acid salt of benzyl
1-(3-(((S)-2-(tert-butoxycarbonyl)aminopropanamido)
methyl)phenyl)-3-(trifluoromethyl)-1H-pyrazol-5-ylcarbamate.
[0442] NMR (CD3OD): 7.51-7.26 (m, 9H), 6.72 (s, 1H), 5.13 (s, 2H),
4.46 (m, 2H), 3.92 (q, J=7.1 Hz, 1H), 1.51 (d, J=7.1 Hz, 3H).
[0443] MS m/z 462 (M+H)
(2S)--N-(3-(5-(2-phenylacetamido)-3-(trifluoromethyl)-1H-pyrazol-1-yl)benz-
yl)-2-aminopropanamide (Example 188)
Example 188
[0444] ##STR212##
Part A: Preparation of tert-butyl
(S)-1-(3-(5-amino-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzylamino)-1-oxop-
ropan-2-ylcarbamate
[0445] 10% Palladium on carbon (6 mg) was added to the solution of
benzyl 1-(3-(((S)-2-(tert-butoxycarbonyl)aminopropanamido)
methyl)phenyl)-3-(trifluoromethyl)-1H-pyrazol-5-ylcarbamate (9 mg,
0.016 mmol) in methanol (1 Ml). The suspension was stirred under
the blanket of hydrogen gas (using balloon) for 4 h. The reaction
mixture was filtered over a pad of celite and the celite was washed
with methanol. The combined methanol layer was concentrated in
vaccuo to afford 6.8 mg of tert-butyl
(S)-1-(3-(5-amino-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzylamino)-1-oxop-
ropan-2-ylcarbamate.
[0446] MS m/z 428.3 (M+H)
Part B: Preparation of tert-butyl
(S)-1-(3-(5-(2-phenylacetamido)-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzy-
lamino)-1-oxopropan-2-ylcarbamate
[0447] 2-phenylacetyl chloride (5 mg, 0.03 mmol) was added to the
solution of tert-butyl
(S)-1-(3-(5-amino-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzylamino)-1-oxop-
ropan-2-ylcarbamate (6 mg, 0.014 mmol) and triethylamine (7 mg,
0.07 mmol) in dichloromethane (1 Ml). The reaction mixture was
stirred at room temperature for 16 h. The reaction was concentrated
in vaccuo and the crude product was re-dissolved in 1 mL of
methanol and loaded onto preparative HPLC column and purified using
conditions as shown below. [0448] Conditions: Column--YMC ODS
(20.times.50 mm) [0449] Solvents--A--90% water--10% methanol--0.1%
TFA [0450] B--10% water--90% methanol--0.1% TFA [0451]
Gradient--25% B to 100% B in 12 min [0452] Retention time: 9.2
min
[0453] MS m/z 546.5 (M+H)
Part B: Preparation of
(2S)--N-(3-(5-(2-phenylacetamido)-3-(trifluoromethyl)-1H-pyrazol-1-yl)ben-
zyl)-2-aminopropanamide trifluoroacetic acid salt
[0454] tert-Butyl
(S)-1-(3-(5-(2-phenylacetamido)-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzy-
lamino)-1-oxopropan-2-ylcarbamate (3 mg) in dichloromethane (0.2
mL) was treated with a trifluoroacetic acid (50 Ul). The reaction
mixture was stirred at room temperature for 30 minutes. The
reaction mixture was concentrated in vaccuo to furnish 2 mg of
trifluoroacetate salt of
(2S)--N-(3-(5-(2-phenylacetamido)-3-(trifluoromethyl)-1H-pyrazol-1-yl)ben-
zyl)-2-aminopropanamide
[0455] NMR (CD3OD): 7.43-7.22 (m, 9H), 6.77 (s, 1H), 4.44 (m, 2H),
3.92 (q, J=7.1 Hz, 1H), 3.62 (s, 1H), 1.51 (d, J=7.1 Hz, 3H).
[0456] MS m/z 446 (M+H)
1-(1-(3-(((S)-2-aminopropanamido)methyl)phenyl)-3-(trifluoromethyl)-1H-pyr-
azol-5-yl)-3-phenylurea (Example 189)
Example 189
[0457] ##STR213##
[0458] Phenyl isocyanate (6 mg, 0.05 mmol) was added to the
solution of tert-butyl
(S)-1-(3-(5-amino-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzylamino)-1-oxop-
ropan-2-ylcarbamate (13 mg, 0.03 mmol) and triethylamine (10 mg,
0.1 mmol) in dichloromethane (0.5 mL). The reaction mixture was
stirred at 45.degree. C. for 24 h. The reaction mixture was diluted
with 5 Ml of dichloromethane and 2 mL water. The organic layer was
washed with water, dried (over Na2SO4) and concentrated. The crude
was purified using silica gel column chromatography (2%
MeOH--CH.sub.2Cl.sub.2) to yield 7 mg of tert-butyl
(S)-1-(3-(5-(3-phenylureido)-3-(trifluoromethyl)-1H-pyrazol-1-yl)benzylam-
ino)-1-oxopropan-2-ylcarbamate
[0459] The product was taken up in a mixture of trifluoroacetic
acid and dichloromethane (50:200 Ul) and stirred at room
temperature for 45 minutes. The reaction mixture was concentrated
in vaccuo to furnish 7 mg of trifluoroacetic acid salt of
1-(1-(3-(((S)-2-aminopropanamido)methyl)phenyl)-3-(trifluoromethyl)-1H-py-
razol-5-yl)-3-phenylurea.
[0460] NMR (CD3OD): 7.61-7.47 (m, 4H), 7.35 (m, 2H), 7.27 (m, 2H),
7.03 (m, 1H), 6.83 (s, 1H), 4.53 (m, 2H), 3.92 (q, J=7.1 Hz, 1H),
1.50 (d, J=7.1 Hz, 3H).
[0461] MS m/z 447.4 (M+H)
3'-[((S)-2-Amino-propionylamino)-methyl]-biphenyl-2-carboxylic acid
benzylamide (Example 190)
Example 190
[0462] ##STR214##
Part A: Preparation of 3'-Aminomethyl-biphenyl-2-carboxylic acid
methyl ester
[0463] Potassium carbonate (276 mg, 2 mmol) was added to the
stirred suspension of 2-Iodo-benzoic acid methyl ester (157 MG, 0.6
mmol), 2-aminomethylphenylboronic acid (94 mg, 0.5 mmol) and Pd
(0)(PPh3)4 (11.6 mg, 0.01 mmol) in a mixture of
toluene-methanol-water (1.5:0.7:0.7 Ml). The reaction mixture was
heated to 900 for 16 h. It was partitioned between water and
dichloromethane (10 mL each). The aqueous layer was re-extracted
with 10 mL of dichloromethane. Combined organic layer was washed
with water; dried (Na2SO4) and concentrated to yield 32 mg of crude
3'-Aminomethyl-biphenyl-2-carboxylic acid methyl ester which was
used as it in the next reaction.
[0464] MS m/z 242.3 (M+H)
Part B: Preparation of
3'-[(2-tert-Butoxycarbonylamino-propionylamino)-methyl]-biphenyl-2-carbox-
ylic acid methyl ester
[0465] Boc-Ala-Osu (40 mg, 0.14 mmol) was added to the solution of
crude 3'-Aminomethyl-biphenyl-2-carboxylic acid methyl ester (29
mg, 0.12 mmol) and triethylamine (20 uL, 0.18 mmol) in
dichloromethane (1 Ml). The reaction mixture was stirred at room
temperature for 16 h. The reaction was diluted with 10 mL of
dichloromethane and 2 mL of water. The organic layer was separated
and washed with water, dried (Na2SO4) and concentrated to yield the
crude which was purified using silica gel chromatography (25%
EtOAc-hexane) to afford 30 mg of
3'-[(2-tert-Butoxycarbonylamino-propionylamino)-methyl]-biphenyl-2-carbox-
ylic acid methyl ester.
[0466] NMR (CDCl3): 7.9 (m, 2H), 7.5 (d, 1H), 7.3 (m, 2H), 7.2 (m,
1H), 6.9 (m, 2H), 4.6 (bs, 2H), 4.19 (m, 1H), 3.6 (s, 3H), 1.4 (s,
9H), 1.3 (d, 3H)
Part B: Preparation of
3'-[(2-tert-Butoxycarbonylamino-propionylamino)-methyl]-biphenyl-2-carbox-
ylic acid
[0467] 1 M aqueous solution of sodium hydroxide (0.36 mL) was added
to the solution of
3'-[(2-tert-Butoxycarbonylamino-propionylamino)-methyl]-biphenyl-2-carbox-
ylic acid methyl ester (30 mg, 0.073 mmol) in methanol (0.5 Ml).
The solution was stirred at room temperature for 16 h and then
concentrated in vaccuo. The residue was acidified with 10% acetic
acid in water. The aqueous layer was diluted with ethyl acetate (5
Ml). The organic layer was separated and washed with water, dried
(Na2SO4) and concentrated to furnish 22 mg of
3'-[(2-tert-Butoxycarbonylamino-propionylamino)-methyl]-biphenyl-2-carbox-
ylic acid
[0468] MS m/z 399 (M+H)
Part C: Preparation of
3'-[((S)-2-Amino-propionylamino)-methyl]-biphenyl-2-carboxylic acid
benzylamide trifluoroacetate salt
[0469] EDCI (15 mg, 0.04 mmol) was added to the solution of
3'-[(2-tert-Butoxycarbonylamino-propionylamino)-methyl]-biphenyl-2-carbox-
ylic acid (10 mg, 0.025 mmol), HOAt (11 mg, 0.04 mmol) and
N,N-diisopropylethylamine (40 uL) in dichloromethane (0.3 Ml).
After stirring at room temperature for 20 minutes, benzyl amine
(5.4 mg, 0.05 mmol) was added and the reaction mixture was stirred
at room temperature for additional 16 h. Diluted with 5 mL
dichloromethane and 1 mL of water. The organic layer was washed
with water, dried (Na2SO4) and concentrated. The crude was
suspended in a mixture of trifluoroacetic acid and dichloromethane
(0.1:0.3 Ml). The reaction mixture was stirred at room temperature
for 30 minutes and then concentrated in vaccuo to afford the crude
which was purified using the conditions below to furnish 10 mg of
3'-[((S)-2-Amino-propionylamino)-methyl]-biphenyl-2-carboxylic acid
benzylamide trifluoroacetate salt [0470] Conditions: Column--YMC
ODS (20.times.50 mm) [0471] Solvents--A--90% water--10%
methanol--0.1% TFA [0472] B--10% water--90% methanol--0.1% TFA
[0473] Gradient--15% B to 100% B in 12 min [0474] Retention time:
9.2 min
[0475] NMR (CD3OD): 7.54-7.20 (m, 11H), 7.02 (m, 2H), 4.46-4.33 (m,
4H), 3.92 (q, J=7.1 Hz, 1H), 1.50 (d, J=7.1 Hz, 3H)
[0476] MS m/z 388.5 (M+H)
(2S)--N-(3-(5-(5-(2-methylbenzo
[d]oxazol-7-yl)-1,3,4-oxadiazol-2-yl)-3-(trifluoromethyl)-1H-pyrazol-1-yl-
)benzyl)-2-aminopropanamide (Example 190)
Example 190
[0477] ##STR215##
Part A: Preparation of 2-methylbenzo[d]oxazole-7-carboxylic
acid
[0478] A vial was charged with 3-amino-2-hydroxybenzoic acid (0.5
g, 3.26 mmol) in triethylorthoacetate (2 mL) and p-toluenesulfonic
acid (20 mg). The reaction mixture was heated for 5 hours at
100.degree. C. The reaction mixture was concentrated in vaccuo and
the residue (0.55 g) was used as such in the next reaction.
[0479] MS m/z 178 (M+H)
Part B: Preparation of
(2S)--N-(3-(5-(5-(2-methylbenzo[d]oxazol-7-yl)-1,3,4-oxadiazol-2-yl)-3-(t-
rifluoromethyl)-1H-pyrazol-1-yl)benzyl)-2-aminopropanamide
[0480] A vial containing 2-methylbenzo[d]oxazole-7-carboxylic acid
(56.5 mg, 0.32 mmol),
1-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-3-(trifluoro-
methyl)-1H-pyrazole-5-carboxylic acid hydrazide (100 mg, 0.21
mmol), PyBOP (165 mg, 0.31 mmol) and triethylamine (74 .mu.L) in
DMF (2 mL) was heated in microwave oven at 120.degree. C. for 20
minutes.
[0481] The reaction mixture was cooled at room temperature and to
which was added diisopropylcarbodidimide (134 mg, 1.06 mmol) and
the reaction was heated to 140.degree. C. for 30 minutes in a
microwave oven. The reaction was diluted with methanol (2 mL) and
directly loaded onto preparative HPLC column. The crude product was
purified using the conditions as shown below to yield tert-butyl
(S)-1-(3-(5-(5-(2-methylbenzo[d]oxazol-7-yl)-1,3,4-oxadiazol-2-yl)-3-(tri-
fluoromethyl)-1H-pyrazol-1-yl)benzylamino)-1-oxopropan-2-ylcarbamate
(10 mg) [0482] Conditions: Column--YMC ODS (20.times.50 mm) [0483]
Solvents--A--90% water--10% methanol--0.1% TFA [0484] B--10%
water--90% methanol--0.1% TFA [0485] Gradient--25% B to 100% B in
12 min [0486] Retention time: 9.6 min
[0487] tert-butyl
(S)-1-(3-(5-(5-(2-methylbenzo[d]oxazol-7-yl)-1,3,4-oxadiazol-2-yl)-3-(tri-
fluoromethyl)-1H-pyrazol-1-yl)benzylamino)-1-oxopropan-2-ylcarbamate
(10 mg) was treated with a mixture of trifluoroacetic acid in
dichloromethane (1:3, 2 Ml) for 2 hours at room temperature. Then
the reaction mixture was concentrated in vacuo to yield
(2S)--N-(3-(5-(5-(2-methylbenzo[d]oxazol-7-yl)-1,3,4-oxadiazol-2-yl)-3-(t-
rifluoromethyl)-1H-pyrazol-1-yl)benzyl)-2-aminopropanamide (6 mg)
as trifluoroacetate salt.
[0488] NMR (CD3OD): 8.2-8 (m, 1H), 7.8 (s, 1H), 7.7 (m, 1H),
7.52-7.4 (m, 2H), 7.3-7.1 (m, 2H), 4.37 (dd, 2H), 3.7 (q, J=7.1 Hz,
1H), 2.6 (s, 3H), 1.50 (d, J=7.1 Hz, 3H)
[0489] MS m/z 512.1 (M+H)
(2S)--N-(3-(5-(3-(benzo
[d]thiazol-7-yl)-1,2,4-oxadiazol-5-yl)-3-(trifluoromethyl)-1H-pyrazol-1-y-
l)benzyl)-2-aminopropanamide (Example 191)
Example 191
[0490] ##STR216##
Part A: Preparation of benzo[d]thiazole-7-carbaldehyde
[0491] Isobutyl chloroformate (715 mg, 5.5 mmol) was added to the
cooled (0.degree. C.) solution of benzo[d]thiazole-7-carboxylic
acid (895 mg, 5 mmol) and N-methylmorpholine (1.2 mL, 11.0 mmol) in
dichloromethane (15 mL). After 15 minutes of stirring,
N-methoxymethanamine hydrochloride (488 mg, 5 mmol) was added and
the reaction mixture was stirred at room temperature for 16 hours.
The reaction mixture was washed with 0.2 M KHSO.sub.4 (2.times.10
mL); followed by water and the combined organic layer was dried
(Na.sub.2SO.sub.4) and concentrated. The product was used as such
in the next reaction.
[0492] Di-isobutyl aluminium hydride (8 mL, 8.0 mmol, 1M soln in
toluene) was added to the cooled solution of crude product as
obtained above (890 mg, 4 mmol) in dry tetrahydrofuran (30 mL)
cooled at -78.degree. C. The reaction mixture was stirred at
-78.degree. C. for 1 hour and then quenched with 1 mL of ethyl
acetate. The reaction mixture was poured into a mixture of ether
and 1M aqueous citric acid. The layers were separated. The aquesous
layer was extracted with ethyl acetate (20 mL). The combined
organic layers were washed with water, dried (Na.sub.2SO.sub.4) and
concentrated. The crude product was purified using silica gel
column chromatography (30% ethyl acetate-hexane) to yield 325 mg of
benzo[d]thiazole-7-carbaldehyde.
[0493] NMR (CDCl.sub.3): 10.7(s, 1H), 9.75 (s, 1H), 8.3 (d, 1H),
7.9 (d, 1H), 7.63 (m, 1H)
[0494] MS m/z 164 (M+H)
Part B: Preparation of benzo[d]thiazole-7-carbonitrile
[0495] A mixture of benzo[d]thiazole-7-carbaldehyde (200 mg, 1.2
mmol) and hydroxylamine hydrochloride (128 mg, 1.8 mmol) in a
mixture of triethylamine and acetonitrile (260 .mu.L: 10 mL) was
heated at 70.degree. C. for 2 hours. Solvent was evaporated under
vacuo and the residue was used as such in the next reaction.
[0496] The crude product was treated with triphenylphosphine (390
mg, 1.5 mmol) and N-chlorosuccinamide (197 MG, 1.5 mmol) in
dichloromethane (5 mL). The reaction mixture was stirred at room
temperature for 16 hours and then concentrated in vacuo. The crude
was purified using silica gel column chromatography (30% ethyl
acetate-hexane) to afford 165 mg of
benzo[d]thiazole-7-carbonitrile.
[0497] NMR (CDCl.sub.3): 9.65 (s, 1H), 8.5 (d, 1H), 7.9 (d, 1H),
7.63 (m, 1H)
Part C: Preparation of
N'-hydroxybenzo[d]thiazole-7-carboxamidine
[0498] A mixture of benzo[d]thiazole-7-carbonitrile (20 mg, 0.13
mmol), hydroxylamine hydrochloride (13 mg, 0.19 mmol) and
triethylamine (26.1 .mu.L, 0.19 mmol) in ethanol (3 mL) was heated
to 70.degree. C. for 2 hours. The solvent was evaporated and the
crude product was purified using preparative HPLC using conditions
as described below to give 29 mg of
N'-hydroxybenzo[d]thiazole-7-carboxamidine [0499] Conditions:
Column--YMC ODS (20.times.50 mm) [0500] Solvents--A--90% water--10%
methanol--0.1% TFA [0501] B--10% water--90% methanol--0.1% TFA
[0502] Gradient--15% B to 100% B in 12 min [0503] Retention time:
8.6 min
[0504] MS m/z 194 (M+H)
Part D: Preparation of
(2S)--N-(3-(5-(3-(benzo[d]thiazol-7-yl)-1,2,4-oxadiazol-5-yl)-3-(trifluor-
omethyl)-1H-pyrazol-1-yl)benzyl)-2-aminopropanamide (Example
191)
[0505] A mixture of
1-(3-(((S)-2-(tert-butoxycarbonyl)propanamido)methyl)phenyl)-3-(trifluoro-
methyl)-1H-pyrazole-5-carboxylic acid (32 mg, 0.07 mmol), HBTU (27
mg, 0.07 mmol), N'-hydroxybenzo[d]thiazole-7-carboxamidine (13.5
mg, 0.07 mmol) and N,N'-diisopropylethylamine (29 .mu.L, 0.17 mmol)
in dry DMF (1 mL) was heated in microwave oven at 190.degree. C.
for 2 minutes. The reaction mixture was diluted with a mixture of
ethyl acetate-water (20:5 mL); the organic layer was separated;
washed with water; dried (Na.sub.2SO.sub.4) and concentrated to
afford a crude.
[0506] The crude was treated with trifluoroacetic acid (0.2 mL) in
dichloromethane (0.8 mL). The reaction mixture was concentrated,
the resulting crude was dissolved in methanol and loaded onto
preparative HPLC. The following conditions were used for the
purification to yield 22 mg of
(2S)--N-(3-(5-(3-(benzo[d]thiazol-7-yl)-1,2,4-oxadiazol-5-yl)-3-(tr-
ifluoromethyl)-1H-pyrazol-1-yl)benzyl)-2-aminopropanamide as
trifluoroacetic acid salt. [0507] Conditions: Column--YMC ODS
(20.times.50 mm) [0508] Solvents--A--90% water--10% methanol--0.1%
TFA [0509] B--10% water--90% methanol--0.1% TFA [0510]
Gradient--15% B to 100% B in 12 min [0511] Retention time: 9.4
min
[0512] NMR (CD3OD): 9.65 (s, 1H), 8.1-8.3 (m, 2H), 7.7-7.9 (m, 3H),
7.1 (m, 3H), 4.4 (dd, 2H), 3.8 (m, 1H), 1.3 (d, 3H)
[0513] MS m/z 514.2 (M+H)
* * * * *